U.S. patent application number 17/191809 was filed with the patent office on 2021-06-24 for method and analysis system for testing a sample.
This patent application is currently assigned to Boehringer Ingelheim Vetmedica GmbH. The applicant listed for this patent is Boehringer Ingelheim Vetmedica GmbH. Invention is credited to Guenter Bruckmann, Erol Meyda, Axel Niemeyer, Harald Pauls.
Application Number | 20210187509 17/191809 |
Document ID | / |
Family ID | 1000005434685 |
Filed Date | 2021-06-24 |
United States Patent
Application |
20210187509 |
Kind Code |
A1 |
Bruckmann; Guenter ; et
al. |
June 24, 2021 |
METHOD AND ANALYSIS SYSTEM FOR TESTING A SAMPLE
Abstract
An analysis system for testing, in particular, a biological
sample, the sample and/or analytes of the sample being
temperature-controlled in advance or denatured immediately before
the test, and/or being heated, the heat being transferred through a
chip of a sensor apparatus.
Inventors: |
Bruckmann; Guenter;
(Wuerselen, DE) ; Meyda; Erol; (Aachen, DE)
; Niemeyer; Axel; (Bielefeld, DE) ; Pauls;
Harald; (Eschweiler, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Boehringer Ingelheim Vetmedica GmbH |
Ingelheim am Rhein |
|
DE |
|
|
Assignee: |
Boehringer Ingelheim Vetmedica
GmbH
Ingelheim am Rhein
DE
|
Family ID: |
1000005434685 |
Appl. No.: |
17/191809 |
Filed: |
March 4, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
15725323 |
Oct 5, 2017 |
10953403 |
|
|
17191809 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C12Q 1/686 20130101;
B01L 2300/1822 20130101; B01L 2300/0883 20130101; B01L 2300/0636
20130101; B01L 2300/0809 20130101; B01L 2300/0645 20130101; B01L
7/52 20130101; B01L 2200/147 20130101; C12Q 1/6825 20130101; B01L
2300/1827 20130101; B01L 3/5027 20130101; B01L 2300/0819 20130101;
C12Q 1/6806 20130101; B01L 2200/10 20130101; B01L 2300/0816
20130101 |
International
Class: |
B01L 7/00 20060101
B01L007/00; B01L 3/00 20060101 B01L003/00; C12Q 1/6806 20060101
C12Q001/6806; C12Q 1/6825 20060101 C12Q001/6825; C12Q 1/686
20060101 C12Q001/686 |
Foreign Application Data
Date |
Code |
Application Number |
Oct 7, 2016 |
EP |
16 020 371.7 |
Claims
1. An analysis system for testing a biological sample, comprising:
at least one receiving cavity for the sample, a plurality of
reaction cavities for forming amplification products of analytes of
the sample, and a sensor apparatus for detecting at least one of
the analytes or the amplification products, wherein the reaction
cavities are fluidically arranged between the at least one
receiving cavity and the sensor apparatus, wherein the sensor
apparatus comprises a sensor compartment fluidically linked to the
plurality of reaction cavities, wherein the sensor apparatus
comprises capture molecules for bonding at least one of the
analytes or the amplification products, and wherein the capture
molecules have different hybridization temperatures at which the at
least one of the analytes or the amplification products bonds to
the corresponding capture molecules.
2. The analysis system according to claim 1, further comprising a
mixing cavity for mixing the sample with at least one reagent,
wherein the mixing cavity is fluidically arranged between the at
least one receiving cavity and the plurality of reaction
cavities.
3. The analysis system according to claim 2, wherein the mixing
cavity is provided with at least one reagent.
4. The analysis system according to claim 1, further comprising a
plurality of groups of reagents for carrying out an amplification
reaction for forming the amplification products.
5. The analysis system according to claim 4, wherein a different
group of the plurality of groups of reagents is assigned to each
reaction cavity of the plurality of reaction cavities.
6. The analysis system according to claim 4, wherein each group of
the plurality of groups of reagents comprises a group of primers
for carrying out the amplification reaction.
7. The analysis system according to claim 6, wherein the groups of
primers differ from one another such that amplification products
with different hybridization temperatures can be formed in the
plurality of reaction cavities.
8. The analysis system according to claim 4, wherein the plurality
of groups of reagents are arranged in the plurality of reaction
cavities.
9. The analysis system according to claim 4, the analysis system
further comprising a plurality of intermediate cavities, wherein
the plurality of groups of reagents are contained in the
intermediate cavities.
10. The analysis system according to claim 9, wherein the
intermediate cavities are at least one of fluidically arranged in
parallel to one another or between the plurality of reaction
cavities and the at least one receiving cavity.
11. The analysis system according to claim 1, wherein the reaction
cavities are fluidically arranged in parallel to one another.
12. The analysis system according to claim 1, further comprising an
intermediate temperature-control cavity for actively
temperature-controlling at least one of the analytes or
amplification products, wherein the intermediate
temperature-control cavity is fluidically arranged between the
plurality of reaction cavities on one side and the sensor apparatus
on another side.
13. The analysis system according to claim 12, further comprising
an intermediate temperature-control apparatus for actively
temperature-controlling the intermediate temperature-control
cavity.
14. The analysis system according to claim 12, wherein the sensor
apparatus is fluidically connected to all the of the reaction
cavities via the intermediate temperature-control cavity.
15. The analysis system according to claim 1, wherein the sensor
apparatus comprises a support and a plurality of electrodes
arranged on the support, wherein a sensor temperature-control
apparatus is provided for directly temperature-controlling the
support.
16. The analysis system according to claim 15, wherein the support
comprises a chip or is formed by a chip.
17. The analysis system according to claim 1, further comprising a
connection apparatus for at least one of electrically or thermally
connecting the sensor apparatus with an analysis device.
18. The analysis system according to claim 17, wherein the
connection apparatus is movable against the sensor apparatus.
19. The analysis system according to claim 17, wherein the
connection apparatus comprises a sensor temperature-control
apparatus for controlling the temperature of the sensor
apparatus.
20. The analysis system according to claim 1, further comprising a
cartridge for receiving the sample and an analysis device for
receiving the cartridge.
21. Analysis system according to claim 20, wherein the cartridge
comprises at least one of the at least one receiving cavity, the
sensor apparatus or the plurality of reaction cavities.
22. The analysis system according to claim 20, wherein the analysis
device comprises at least one of a reaction temperature-control
apparatus assigned to the plurality of reaction cavities, an
intermediate temperature-control apparatus assigned to an
intermediate temperature-control cavity, a sensor
temperature-control apparatus for temperature-controlling the
sensor apparatus or a connection apparatus for at least one of
electrically or thermally connecting the sensor apparatus to the
cartridge.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a division of U.S. patent application
Ser. No. 15/725,323 filed Oct. 5, 2017, which claims the benefit of
priority to European Patent Application No. 16 020 371.7 filed Oct.
7, 2016, the contents of which are incorporated herein by reference
in their entirety.
BACKGROUND OF THE INVENTION
Field of the Invention
[0002] The present invention relates to an analysis system and
method in which analytes of a sample are pre-treated and/or
amplification products produced from analytes of the sample in a
reaction cavity.
[0003] Preferably, the present invention deals with analysing and
testing a sample, in particular from a human or animal,
particularly preferably for analytics and diagnostics, for example
with regard to the presence of diseases and/or pathogens and/or for
determining blood counts, antibodies, hormones, steroids or the
like. Therefore, the present invention is in particular within the
field of bioanalytics. A food sample, environmental sample or
another sample may optionally also be tested, in particular for
environmental analytics or food safety and/or for detecting other
substances.
[0004] In particular, by means of the present invention, at least
one analyte (target analyte) of a sample, preferably a nucleic-acid
product, such as a particular nucleic-acid sequence, can be
determined, identified or detected. In particular, the sample can
be tested for qualitatively or quantitatively determining at least
one analyte, for example in order for it to be possible to detect a
disease and/or pathogen.
[0005] The present invention deals in particular with what are
known as point-of-care systems, i.e., with systems, devices and
other apparatuses, and deals with methods for carrying out tests on
a sample at the sampling site and/or independently or away from a
central laboratory or the like.
Description of Related Art
[0006] U.S. Pat. No. 5,096,669 discloses a point-of-care system for
testing a biological sample, in particular a blood sample. The
system comprises a single-use cartridge and an analysis device.
Once the sample has been received, the cartridge is inserted into
the analysis device in order to carry out the test. The cartridge
comprises a microfluidic system and a sensor apparatus comprising
electrodes, the apparatus being calibrated by means of a
calibration liquid and then being used to test the sample.
[0007] Furthermore, International Patent Application Publication WO
2006/125767 A1 and U.S. Patent Application Publication 2016/0047832
A1 disclose a point-of-care system for integrated and automated DNA
or protein analysis, comprising a single-use cartridge and an
analysis device for fully automatically processing and evaluating
molecular-diagnostic analyses using the single-use cartridge. The
cartridge is designed to receive a sample, in particular blood, and
in particular allows cell disruption, PCR and detection of PCR
amplification products, which are bonded to capture molecules and
provided with a label enzyme, in order for it to be possible to
detect bonded PCR amplification products or nucleic sequences as
target analytes in what is known as a redox cycling process. The
temperature in the PCR chamber can be manipulated by associated
Peltier elements.
[0008] U.S. Patent Application Publication 2009/0227476 A1
discloses a biological assay apparatus which can perform PCR in a
PCR thermal cycling chamber and, thereafter, can detect nucleic
acids in a microarray. The PCR thermal cycling chamber can be
separately heated or cooled.
SUMMARY OF THE INVENTION
[0009] The problem addressed by the present invention is to provide
an improved analysis system and an improved method for testing an
in particular biological sample, which allow or facilitate testing
of the sample that is efficient, reliable, rapid and/or as precise
as possible.
[0010] The above problem is solved by an analysis system and a
method as described herein.
[0011] The proposed analysis system for testing, in particular, a
biological sample preferably comprises a receiving cavity for
receiving the sample and/or at least one reaction cavity for
amplifying analytes of the sample and/or for forming amplification
products from the analytes of the sample.
[0012] Preferably, the analysis system comprises a sensor apparatus
that is in particular separate from the receiving cavity and/or
reaction cavity, and/or is spaced apart from the receiving cavity
and/or reaction cavity, in order to in particular electrochemically
detect the analytes and/or amplification products that are
preferably bonded to capture molecules of the sensor apparatus.
[0013] According to a first aspect of the present invention, the
analysis system comprises an intermediate temperature-control
cavity for actively temperature-controlling, in particular heating,
the amplified analytes and/or amplification products, the
intermediate temperature-control cavity being arranged between the
receiving cavity and/or reaction cavity on one side and the sensor
apparatus on the other side, and/or the sensor apparatus being
fluidically connected to the receiving cavity and/or reaction
cavity by the intermediate temperature-control cavity.
[0014] Advantageously, by means of the intermediate
temperature-control cavity it is possible to prevent or reduce
undesired hybridization of the analytes and/or amplification
products before they are fed to the sensor apparatus, and/or to
separate or denature analytes and/or amplification products that
are bonded to one another, in particular such that the number of
analytes and/or amplification products that are unbonded or
available for hybridization to the corresponding capture molecules
is maximised or at least increased. In this way, the yield of
amplification products bonded to capture molecules, and thus the
efficiency of the test, is increased.
[0015] In particular, by means of the intermediate
temperature-control cavity it is possible to denature and/or
temperature-control the analytes and/or amplification products
amplified in the reaction cavity before they are fed to the sensor
apparatus, preferably such that the analytes and/or amplification
products can be fed to the sensor apparatus in the denatured state
and/or the state in which they are temperature-controlled in
advance. Therefore, denaturing of the analytes and/or amplification
products in or on the sensor apparatus can be omitted, and/or the
required temperature control of the amplification products in or on
the sensor apparatus can be reduced, as a result of which the
efficiency of the test is increased.
[0016] According to another aspect of the present invention, which
can also be implemented independently, the analysis system
comprises, in particular in addition to the intermediate
temperature-control cavity, a sensor temperature-control apparatus
for in particular directly temperature-controlling the sensor
apparatus and/or for temperature-controlling the capture molecules
and analytes and/or amplification products in the sensor apparatus,
in particular so that heat is transferred from the sensor
temperature-control apparatus through the sensor apparatus to the
capture molecules, analytes and/or amplification products or vice
versa.
[0017] Particularly preferably, different temperatures of the
capture molecules and analytes and/or amplification products in or
on the sensor apparatus can be set or reacted by means of the
sensor temperature-control apparatus, preferably such that
(different) analytes and/or amplification products can be bonded to
the corresponding capture molecules at different hybridization
temperatures. Advantageously, the efficiency and/or specificity of
the test and/or detection is thus increased.
[0018] Preferably, the sensor apparatus comprises a support, in
particular a chip as the support, a plurality of electrodes
arranged on the support and/or a sensor compartment, the sensor
temperature-control apparatus preferably being designed for in
particular directly temperature-controlling the support and/or the
sensor compartment, in particular so that heat is transferred from
the sensor temperature-control apparatus through the support to the
sensor compartment or vice versa, and/or the sensor
temperature-control apparatus, in the operating state, resting on
or against the support or the back thereof, in particular in a
planar manner and/or centrally, in order to temperature-control the
sensor compartment from outside and/or in particular with heat
being transferred through the support.
[0019] By means of the sensor temperature-control apparatus, it is
possible to achieve different temperatures or temperature curves or
temperature profiles in or on the sensor apparatus and/or in the
sensor compartment, and/or to adapt the temperature control of the
support and/or sensor compartment for (optimally) hybridizing the
amplification products to the capture molecules.
[0020] The proposed analysis system is in particular portable,
mobile and/or is a point-of-care system and/or can be used in
particular at the sampling site and/or away from a central
laboratory.
[0021] The analysis system preferably comprises an analysis device
and/or at least one cartridge for testing the sample.
[0022] The term "analysis device" is preferably understood to mean
an instrument which is in particular mobile and/or can be used on
site, and/or which is designed to chemically, biologically and/or
physically test and/or analyse a sample or a component thereof,
preferably in and/or by means of a cartridge. In particular, the
analysis device controls the testing of the sample in the
cartridge.
[0023] Particularly preferably, the analysis device is designed to
receive the cartridge or to connect said cartridge.
[0024] The term "cartridge" is preferably understood to mean a
structural apparatus or unit designed to receive, to store, to
physically, chemically and/or biologically treat and/or prepare
and/or to measure a sample, preferably in order to make it possible
to detect, identify or determine at least one analyte of the
sample.
[0025] A cartridge within the meaning of the present invention
preferably comprises a fluid system having a plurality of channels,
cavities and/or valves for controlling the flow through the
channels and/or cavities.
[0026] In particular, within the meaning of the present invention,
a cartridge is designed to be at least substantially planar, flat
and/or card-like, in particular is designed as a (micro)fluidic
card and/or is designed as a main body or container that can
preferably be closed and/or said cartridge can be inserted and/or
plugged into a proposed analysis device when it contains the
sample.
[0027] The proposed method for testing an in particular biological
sample provides for actively temperature-controlling, in particular
heating, the analytes of the sample and/or the amplification
products, which are preferably amplified by means of PCR, between
the reaction cavity and the sensor apparatus and/or after
amplification/copying and immediately before hybridization to
capture molecules, particularly preferably in order to denature the
analytes and/or amplification products, and/or prevent or reduce
undesired hybridization of the analytes and/or amplification
products to one another.
[0028] Preferably, different analytes and/or amplification products
or groups thereof are initially, in particular simultaneously
and/or in parallel, produced by means of an amplification reaction,
in particular PCR, in preferably different PCR chambers and/or
reaction cavities, and are then bonded to the capture molecules in
succession at different hybridization temperatures.
[0029] In particular, it is provided that a first, second and
optional third group of amplification products are produced in
different reaction cavities. It may however also be provided that
the analytes are amplified by means of an amplification reaction,
in particular PCR, in a common PCR chamber or reaction cavity,
and/or that the amplification products are produced in a common
reaction cavity.
[0030] Preferably, a plurality of amplification reactions, in
particular PCRs, run simultaneously, in parallel or independently
from one another during the test.
[0031] Preferably, different amplification reactions, in particular
PCRs with different primers, are provided or carried out.
[0032] Within the meaning of the present invention, amplification
reactions are in particular molecular-biological reactions in which
an analyte is amplified/copied and/or in which amplification
products, in particular nucleic-acid products, of an analyte are
produced. Particularly preferably, PCRs are amplification reactions
within the meaning of the present invention.
[0033] "PCR" stands for polymerase chain reaction and is a
molecular-biological method by means of which certain analytes, in
particular portions of RNA or DNA, of a sample are amplified,
preferably in several cycles, using polymerases or enzymes, in
particular in order to then test and/or detect the amplification
products or nucleic-acid products. If RNA is intended to be tested
and/or amplified, before the PCR is carried out, a cDNA is produced
starting from the RNA, in particular using reverse transcriptase.
The cDNA is used as a template for the subsequent PCR.
[0034] Preferably, during a PCR, a sample is first denatured by the
addition of heat in order to separate the strands of DNA or cDNA.
Preferably, primers or nucleotides are then deposited on the
separated single strands of DNA or cDNA, and a desired DNA or cDNA
sequence is replicated by means of polymerase and/or the missing
strand is replaced by means of polymerase. This process is
preferably repeated in a plurality of cycles until the desired
quantity of the DNA or cDNA sequence is available.
[0035] For the PCR, marker primers are preferably used, i.e.,
primers which (additionally) produce a marker or a label, in
particular biotin, on the amplified analyte. This allows or
facilitates detection. Preferably, the primers used are
biotinylated and/or comprise or form in particular covalently
bonded biotin as the label.
[0036] It is proposed that the analytes and/or amplification
products are actively temperature-controlled, preferably in advance
and/or before being temperature-controlled (again) in the sensor
apparatus, preferably (pre-)heated, preferably in an intermediate
temperature-control cavity and/or by means of an intermediate
temperature-control apparatus, after leaving the reaction cavity
and/or after the PCR is carried out and/or immediately before being
fed to the sensor apparatus, in particular in order to separate any
potentially double-stranded analytes and/or amplification products
into single strands.
[0037] Preferably, the analytes and/or amplification products are
then subsequently and/or again temperature-controlled, in
particular after being temperature-controlled in the intermediate
temperature-control cavity, and/or brought to the corresponding
hybridization temperature, in particular in or on the sensor
apparatus and/or by means of a sensor temperature-control
apparatus, preferably in order to hybridize the amplification
products to the corresponding capture molecules.
[0038] Preferably, the sensor compartment, or the capture
molecules, analytes and/or amplification products is/are actively
temperature-controlled and/or brought to the corresponding
hybridization temperature, in particular with heat being
transferred through a support, in particular a chip, of the sensor
apparatus.
[0039] The support is preferably both electrically and thermally
contacted on the back, and/or is connected and/or coupled to the
analysis device. This allows a particularly compact design.
[0040] The hybridization temperature is preferably the (average)
temperature at which an (amplified) analyte, in particular portions
of RNA or DNA, and/or an amplification product is bonded to
corresponding capture molecules and/or is hybridized to
corresponding capture molecules.
[0041] The optimal hybridization temperature is preferably the
temperature at which the number of amplification products bonded to
corresponding capture molecules is maximised and/or the number of
amplification products bonded to one another is minimised.
[0042] Preferably, the (optimal) hybridization temperature varies
for different analytes and/or amplification products.
[0043] A group of different analytes and/or amplification products
preferably only includes, at least substantially, analytes and/or
amplification products having similar (optimal) hybridization
temperatures. Therefore, this results in an average and/or optimal
hybridization temperature of the group or a temperature range of
(optimal) hybridization temperatures. At this temperature or in
this temperature range of the group--both also referred to as
"group temperature" for short--the total number of analytes and/or
amplification products in this group that are bonded to the capture
molecules is (likely to be) maximal. The temperature range is
preferably less than 8.degree. C., in particular less than
5.degree. C.
[0044] Preferably, different groups having different group
temperatures are formed. The group temperatures preferably differ
or are spaced apart by at least 2.degree. C., in particular by more
than 3.degree. C.
[0045] In particular, the group temperature of a first group is
greater than the group temperature of a second group.
[0046] Preferably, the (optimal) hybridization temperature varies
depending on the GC content of the DNA or cDNA, the length of the
DNA or cDNA, the melting point or melting temperature of the DNA or
cDNA sequence and/or the conditioning or salt concentration of the
solvent, ambient medium and/or buffer.
[0047] The melting point or melting temperature is preferably the
temperature at which or from which the DNA or cDNA denatures and/or
the strands of double-stranded DNA or cDNA are separated from one
another. The melting point or melting temperature is preferably
dependent on the GC content of the DNA or cDNA, the length of the
DNA or cDNA, and/or the conditioning or salt concentration of the
solvent, ambient medium and/or buffer. Preferably, the melting
point or melting temperature is at least 85.degree. C. or
90.degree. C., particularly preferably 92.degree. C. or 94.degree.
C., and/or at most 99.degree. C. or 98.degree. C., particularly
preferably at most 97.degree. C. or 96.degree. C.
[0048] Preferably, the hybridization temperature is lower than the
melting point or melting temperature, preferably by at least
2.degree. C. or 5.degree. C., particularly preferably 8.degree. C.
or 10.degree. C., in particular 15.degree. C. or 20.degree. C. or
more.
[0049] The capture molecules of the sensor apparatus are in
particular oligonucleotide probes, which are preferably immobilised
on the sensor, sensor array and/or electrodes by a spacer, in
particular a C6 spacer. The formation of structures that disrupt
hybridization, e.g., hairpin structures, can be prevented by the
preferred bonding of the capture molecules by spacers.
[0050] The analytes and/or amplification products bonded at
different hybridization temperatures are preferably detected in a
single or common detection process.
[0051] Particularly preferably, the sensor apparatus is only used a
single time for a process for detecting said analytes and/or
amplification products, electrochemical determination preferably
taking place in particular simultaneously for all the bonded
amplification products. This allows very rapid and efficient
testing.
[0052] In particular, different analytes and/or amplification
products of different analytes can be very efficiently bonded by
hybridization temperatures in succession, to preferably immobilised
capture molecules, particularly preferably on or in a sensor
apparatus, in order for it to be possible to measure and/or
determine or detect a particularly large number of different
amplification products at the same time, in particular in a single
or common detection process.
[0053] In the context of the present invention, it is thus possible
to test analytes and/or amplification products that are produced
and/or amplified in parallel and have different hybridization
temperatures in a single detection process and at the same time
with high specificity.
[0054] Preferably, the bonded analytes and/or amplification
products are detected by feeding detector molecules to the sensor
apparatus.
[0055] Within the meaning of the present invention, the term
"detector molecules" is preferably understood to mean molecules
that bond specifically to the marker or label of the primers used
to amplify the analytes and/or analytes or amplification products
provided therewith, and thus allow the detection thereof.
[0056] In particular, the detector molecules may be enzyme
conjugates and/or immunoconjugates, which bond specifically to the
marker or label, in particular biotin, and comprise a reporter
enzyme for converting a substrate. In the context of the present
invention, the detector molecules are preferably based on
streptavidin, which has a high affinity for biotin, and/or alkaline
phosphatase, which can convert non-reactive phosphate monoesters to
electrochemically active molecules and phosphate.
[0057] Preferably, a detection system is used, where the label is
based on biotin and where the detector molecules are based on
streptavidin/alkaline phosphatase. However, other detector
molecules can also be used.
[0058] The above-mentioned aspects and features of the present
invention and the aspects and features of the present invention
that will become apparent from the following description can in
principle be implemented independently from one another, but also
in any combination or order.
[0059] Other aspects, advantages, features and properties of the
present invention will become apparent from the following
description of a preferred embodiment with reference to the
accompanying drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0060] FIG. 1 is a schematic section through a proposed analysis
system or analysis device comprising a proposed cartridge received
therein;
[0061] FIG. 2 is a schematic view of the cartridge;
[0062] FIG. 3 is a schematic front view of a proposed sensor
apparatus of the analysis system and/or cartridge;
[0063] FIG. 4 is an enlarged detail from FIG. 3 illustrating a
sensor field of the sensor apparatus;
[0064] FIG. 5 is a schematic rear view of the sensor apparatus;
[0065] FIG. 6 is a schematic sectional view of the sensor
apparatus; and
[0066] FIG. 7 is a schematic curve or profile for the temperature
of the sample and/or of amplification products as a function of the
position in the cartridge.
DETAILED DESCRIPTION OF THE INVENTION
[0067] In the figures, which are only schematic and sometimes not
to scale, the same reference signs are used for the same or similar
parts and components, corresponding or comparable properties and
advantages being achieved even if these are not repeatedly
described.
[0068] FIG. 1 is a highly schematic view of a proposed analysis
system 1 and analysis device 200 for testing an in particular
biological sample P, preferably by means of or in an apparatus or
cartridge 100.
[0069] FIG. 2 is a schematic view of a preferred embodiment of the
proposed apparatus or cartridge 100 for testing the sample P. The
apparatus or cartridge 100 in particular forms a handheld unit, and
in the following is merely referred to as a cartridge.
[0070] The term "sample" is preferably understood to mean the
sample material to be tested, which is in particular taken from a
human or animal. In particular, within the meaning of the present
invention, a sample is a fluid, such as saliva, blood, urine or
another liquid, preferably from a human or animal, or a component
thereof. Within the meaning of the present invention, a sample may
be pre-treated or prepared if necessary, or may come directly from
a human or animal or the like, for example. A food sample,
environmental sample or another sample may optionally also be
tested, in particular for environmental analytics, food safety
and/or for detecting other substances, preferably natural
substances, but also biological or chemical warfare agents, poisons
or the like.
[0071] Preferably, the analysis system 1 or analysis device 200
controls the testing of the sample P in particular in or on the
cartridge 100 and/or is used to evaluate the testing or the
collection, processing and/or storage of measured values from the
test.
[0072] By means of the proposed analysis system 1 or analysis
device 200 or by means of the cartridge 100 and/or using the
proposed method for testing the sample P, preferably an analyte A
of the sample P, in particular a nucleic-acid product, such as a
certain nucleic-acid sequence, or particularly preferably a
plurality of analytes A of the sample P, can be determined,
identified or detected. Said analytes are in particular detected
and/or measured not only qualitatively, but particularly preferably
also quantitatively.
[0073] Therefore, the sample P can in particular be tested for
qualitatively or quantitatively determining at least one analyte A,
for example in order for it to be possible to detect a disease
and/or pathogen or to determine other values, which are important
for diagnostics, for example.
[0074] Particularly preferably, a molecular-biological test is made
possible by means of the analysis system 1 and/or analysis device
200 and/or by means of the cartridge 100.
[0075] Particularly preferably, a molecular and/or PCR assay, in
particular for detecting DNA and/or RNA, i.e., nucleic-acid
products and/or sequences, is made possible and/or carried out.
[0076] Preferably, the sample P or individual components of the
sample P or analytes A can be amplified if necessary, in particular
by means of PCR, and tested, identified or detected in the analysis
system 1, analysis device 200 and/or in the cartridge 100.
Preferably, amplification products V of the analyte A or analytes A
are thus produced.
[0077] The analytes A and/or amplification products V of the sample
P, in particular the nucleic-acid products, which are amplified in
particular by means of PCR, in particular have a length of at least
20 or 50, particularly preferably 80 or 100, and/or at most 300 or
280, particularly preferably 250 or 220, nucleotides. However, it
may also be provided that shorter or longer amplification products
V are produced in particular by means of PCR.
[0078] In the following, further details are first given on a
preferred construction of the cartridge 100, with features of the
cartridge 100 preferably also directly representing features of the
analysis system 1, in particular even without any further explicit
explanation.
[0079] The cartridge 100 is preferably at least substantially
planar, flat and/or plate-shaped and/or card-like.
[0080] The cartridge 100 preferably comprises an in particular at
least substantially flat, planar, plate-shaped and/or card-like
main body 101, the main body 101 in particular being made of and/or
injection-moulded from plastics material, particularly preferably
polypropylene.
[0081] The cartridge 100 preferably comprises at least one film or
cover 102 for covering the main body 101 and/or cavities and/or
channels formed therein at least in part, in particular on the
front 100A, and/or for forming valves or the like, as shown by
dashed lines in FIG. 2.
[0082] The analysis system 1 or cartridge 100 or the main body 101
thereof, in particular together with the cover 102, preferably
forms and/or comprises a fluidic system 103, referred to in the
following as the fluid system 103.
[0083] The cartridge 100 and/or the fluid system 103 thereof is
preferably at least substantially vertically oriented in the
operating position and/or during the test, in particular in the
analysis device 200, as shown schematically in FIG. 1. In
particular, the main plane or surface extension of the cartridge
100 thus extends at least substantially vertically in the operating
position.
[0084] The cartridge 100 and/or the fluid system 103 preferably
comprises a plurality of cavities, in particular at least one
receiving cavity 104, at least one metering cavity 105, at least
one intermediate cavity 106, at least one mixing cavity 107, at
least one storage cavity 108, at least one reaction cavity 109, at
least one intermediate temperature-control cavity 110 and/or at
least one collection cavity 111, as shown in FIG. 1.
[0085] The cartridge 100 and/or the fluid system 103 also
preferably comprises at least one pump apparatus 112 and/or at
least one sensor apparatus 113.
[0086] Some, most or all of the cavities are preferably formed by
chambers and/or channels or other depressions in the cartridge 100
and/or the main body 101, and particularly preferably are covered
or closed by the film or cover 102. However, other structural
solutions are also possible.
[0087] In the example shown, the cartridge 100 or the fluid system
103 preferably comprises two metering cavities 105A and 105B, a
plurality of intermediate cavities 106A to 106G, a plurality of
storage cavities 108A to 108E and/or a plurality of reaction
cavities 109, which can preferably be loaded independently from one
another, in particular a first reaction cavity 109A, a second
reaction cavity 109B and an optional third reaction cavity 109C, as
can be seen in FIG. 2.
[0088] The reaction cavity/cavities 109 is/are used in particular
to carry out an amplification reaction, in particular PCR, or
several, preferably different, amplification reactions, in
particular PCRs. It is preferable to carry out several, preferably
different, PCRs, i.e., PCRs having different primer combinations or
primer pairs, in parallel and/or independently and/or in different
reaction cavities 109.
[0089] The amplification products V and/or other portions of the
sample P forming in the one or more reaction cavities 109 can be
conducted or fed to the connected sensor apparatus 113, in
particular by means of the pump apparatus 112.
[0090] The sensor apparatus 113 is used in particular for
detecting, particularly preferably qualitatively and/or
quantitatively determining, the analyte A or analytes A of the
sample P, in this case particularly preferably the amplification
products V of the analytes A. Alternatively or additionally,
however, other values may also be collected or determined.
[0091] In particular, the pump apparatus 112 comprises or forms a
tube-like or bead-like raised portion, in particular by means of
the film or cover 102, particularly preferably on the back of the
cartridge, as shown schematically in FIG. 1.
[0092] The cartridge 100, the main body 101 and/or the fluid system
103 preferably comprise a plurality of channels 114 and/or valves
115, as shown in FIG. 2.
[0093] By means of the channels 114 and/or valves 115, the cavities
104 to 111, the pump apparatus 112 and/or the sensor apparatus 113
can be temporarily and/or permanently connected and/or separated
from one another, as required and/or optionally or selectively, in
particular such that they are controlled by the analysis system 1
or the analysis device 200.
[0094] The cavities 104 to 111 are preferably each fluidically
linked by a plurality of channels 114. Particularly preferably,
each cavity is linked or connected by at least two associated
channels 114, in order to make it possible for fluid to fill, flow
through and/or drain from the respective cavities as required.
[0095] The fluid transport or the fluid system 103 is preferably
not based on capillary forces, or is not exclusively based on said
forces, but in particular is essentially based on the effects of
gravity and/or pumping forces and/or compressive forces and/or
suction forces that arise, which are particularly preferably
generated by the pump or pump apparatus 112. In this case, the
flows of fluid or the fluid transport and the metering are
controlled by accordingly opening and closing the valves 115 and/or
by accordingly operating the pump or pump apparatus 112, in
particular by means of a pump drive 202 of the analysis device
200.
[0096] Preferably, each of the cavities 104 to 110 has an inlet at
the top and an outlet at the bottom in the operating position.
Therefore, if required, only liquid from the respective cavities
can be removed via the outlet.
[0097] In particular, the liquid-containing cavities, particularly
preferably the storage cavity/cavities 108, the mixing cavity 107
and/or the receiving cavity 104, are each dimensioned such that,
when said cavities are filled with liquid, bubbles of gas or air
that may potentially form rise upwards in the operating position,
such that the liquid collects above the outlet without bubbles.
However, other solutions are also possible here.
[0098] Preferably, at least one valve 115 is assigned to each
cavity, the pump apparatus 112 and/or the sensor apparatus 113
and/or is arranged upstream of the respective inlets and/or
downstream of the respective outlets.
[0099] Preferably, the cavities 104 to 111 or sequences of cavities
104 to 111, through which fluid flows in series or in succession
for example, can be selectively released and/or fluid can
selectively flow therethrough by the assigned valves 115 being
actuated, and/or said cavities can be fluidically connected to the
fluid system 103 and/or to other cavities.
[0100] In particular, the valves 115 are formed by the main body
101 and the film or cover 102 and/or are formed in another manner,
for example by additional layers, depressions or the like.
[0101] Particularly preferably, one or more valves 115A are
provided which are preferably tightly closed initially or when in
storage, particularly preferably in order to seal liquids or liquid
reagents F, located in the storage cavities 108, and/or the fluid
system 103 from the open receiving cavity 104 in a storage-stable
manner.
[0102] Preferably, an initially closed valve 115A is arranged
upstream and downstream of each storage cavity 108. Said valves are
preferably only opened, in particular automatically, when the
cartridge 100 is actually being used and/or while inserting the
cartridge 100 into the analysis device 200.
[0103] A plurality of valves 115A, in particular three valves in
this case, are preferably assigned to the receiving cavity 104 when
an optional intermediate connection 104D is provided in addition to
an inlet 104B and an outlet 104C, for example in order for it to be
possible to optionally discharge or remove a supernatant of the
sample P, such as blood serum or the like. Depending on the use, in
addition to the valve 115A on the inlet 104B, then preferably only
the valve 115A either at the outlet 104C or at the intermediate
connection 104D is opened.
[0104] The valves 115A assigned to the receiving cavity 104 seal
the fluid system 103 and/or the cartridge 100 in particular
fluidically and/or in a gas-tight manner until the sample P is
inserted and the receiving cavity 104 or a connection 104A of the
receiving cavity 104 is closed.
[0105] As an alternative or in addition to the valves 115A (which
are initially closed), one or more valves 115B are preferably
provided which are not closed in a storage-stable manner and/or
which are open initially and/or which can be closed by actuation.
These valves are used in particular to control the flows of fluid
during the test.
[0106] The cartridge 100 is preferably designed as a microfluidic
card and/or the fluid system 103 is preferably designed as a
microfluidic system. In the present invention, the term
"microfluidic" is preferably understood to mean that the respective
volumes of individual cavities, some of the cavities or all of the
cavities 104 to 111 and/or channels 114 are, separately or
cumulatively, less than 5 ml or 2 ml, particularly preferably less
than 1 ml or 800 .mu.l in particular less than 600 .mu.l or 300
.mu.l, more particularly preferably less than 200 .mu.l or 100
.mu.l.
[0107] Particularly preferably, a sample P having a maximum volume
of 5 ml, 2 ml or 1 ml can be introduced into the cartridge 100
and/or the fluid system 103, in particular the receiving cavity
104.
[0108] Reagents and liquids which are preferably introduced or
provided before the test in liquid form as liquids or liquid
reagents F and/or in dry form as dry reagents S are required for
testing the sample P, as shown in the schematic view according to
FIG. 2.
[0109] Furthermore, other liquids F, in particular in the form of a
wash buffer, solvent for dry reagents S and/or a substrate SU, for
example in order to form detection molecules and/or a redox system,
are also preferably required for the test, the detection process
and/or for other purposes and are in particular provided in the
cartridge 100, i.e., are likewise introduced before use, in
particular before delivery. At some points in the following, a
distinction is not made between liquid reagents and other liquids,
and therefore the respective explanations are accordingly also
mutually applicable.
[0110] The analysis system 1 or the cartridge 100 preferably
contains all the reagents and liquids required for carrying out one
or more amplification reactions or PCRs and/or for carrying out the
test, and therefore, particularly preferably, it is only necessary
to receive the optionally pre-treated sample P.
[0111] The cartridge 100 and/or the fluid system 103 preferably
comprises a bypass 114A that can optionally be used, in order for
it to be possible, if necessary, to conduct or convey the sample P
or components thereof past the reaction cavities 109 and, by
bypassing the optional intermediate temperature-control cavity 110,
also directly to the sensor apparatus 113, and/or in order for it
to be possible to convey or pump liquids or liquid reagents F2-F5
out of the storage cavities 108B-108E into the sensor apparatus
113, in particular in the opposite direction to the analytes A
and/or amplification products V, when the bypass 114A is open, more
specifically when the valve 115B of the bypass 114A is open.
[0112] The cartridge 100 or the fluid system 103 or the channels
114 preferably comprise sensor portions 116 or other apparatuses
for detecting liquid fronts and/or flows of fluid.
[0113] It is noted that various components, such as the channels
114, the valves 115, in particular the valves 115A that are
initially closed and the valves 115B that are initially open, and
the sensor portions 116 in FIG. 2 are, for reasons of clarity, only
labelled in some cases, but the same symbols are used in FIG. 2 for
each of these components.
[0114] The collection cavity 111 is preferably used for receiving
excess or used reagents and liquids and volumes of the sample. It
is preferably given appropriate large dimensions and/or is only
provided with inputs or inlets, in particular such that liquids
cannot be removed or pumped out again in the operating
position.
[0115] The receiving cavity 104 preferably comprises a connection
104A for introducing the sample P. After the sample P is introduced
into the receiving cavity 104, said cavity and/or the connection
104A is closed.
[0116] The cartridge 100 can then be inserted into the proposed
analysis device 200 and/or received thereby, as shown in FIG. 1, in
order to test the sample P. Alternatively, the sample P could also
be fed in later.
[0117] FIG. 1 shows the analysis system 1 in a ready-to-use state
for carrying out a test on the sample P received in the cartridge
100. In this state, the cartridge 100 is therefore linked to,
received by and/or inserted into the analysis device 200.
[0118] In the following, some features and aspects of the analysis
device 200 are first explained in greater detail. The features and
aspects relating to said device are preferably also directly
features and aspects of the proposed analysis system 1, in
particular even without any further explicit explanation.
[0119] The analysis system 1 or analysis device 200 preferably
comprises a mount or receptacle 201 for mounting and/or receiving
the cartridge 100.
[0120] Preferably, the cartridge 100 is fluidically, in particular
hydraulically, separated or isolated from the analysis device 200.
In particular, the cartridge 100 forms a preferably independent and
in particular closed fluidic and/or hydraulic system 103 for the
sample P and the reagents and other liquids.
[0121] Preferably, the analysis device 200 is designed to actuate
the pump apparatus 112 and/or valves 115, to have a thermal effect
and/or to detect measured data, in particular by means of the
sensor apparatus 113 and/or sensor portions 116.
[0122] The analysis system 1 or analysis device 200 preferably
comprises a pump drive 202, the pump drive 202 in particular being
designed for mechanically actuating the pump apparatus 112.
[0123] Preferably, a head of the pump drive 202 can be rotated in
order to rotationally axially depress the preferably bead-like
raised portion of the pump apparatus 112. Particularly preferably,
the pump drive 202 and pump apparatus 112 together form a pump, in
particular in the manner of a hose pump or peristaltic pump and/or
a metering pump, for the fluid system 103 and/or the cartridge
100.
[0124] Particularly preferably, the pump is constructed as
described in German Patent DE 10 2011 015 184 B4 and corresponding
U.S. Pat. No. 8,950,424. However, other structural solutions are
also possible.
[0125] Preferably, the capacity and/or discharge rate of the pump
can be controlled and/or the conveying direction of the pump and/or
pump drive 202 can be switched. Preferably, fluid can thus be
pumped forwards or backwards as desired.
[0126] The analysis system 1 or analysis device 200 preferably
comprises a connection apparatus 203 for in particular electrically
and/or thermally connecting the cartridge 100 and/or the sensor
apparatus 113.
[0127] As shown in FIG. 1, the connection apparatus 203 preferably
comprises a plurality of electrical contact elements 203A, the
cartridge 100, in particular the sensor apparatus 113, preferably
being electrically connected or connectable to the analysis device
200 by the contact elements 203A.
[0128] The analysis system 1 or analysis device 200 preferably
comprises one or more temperature-control apparatuses 204, in
particular heating elements or Peltier elements, for
temperature-controlling the cartridge 100 and/or having a thermal
effect on the cartridge 100, in particular for heating and/or
cooling.
[0129] Individual temperature-control apparatuses 204, some of
these apparatuses or all of these apparatuses can preferably be
positioned against the cartridge 100, the main body 101, the cover
102, the sensor apparatus 113 and/or individual cavities and/or can
be thermally coupled thereto and/or can be integrated therein
and/or in particular can be operated or controlled electrically by
the analysis device 200. In the example shown, in particular the
temperature-control apparatuses 204A, 204B and/or 204C are
provided.
[0130] Preferably, the temperature-control apparatus 204A, referred
to in the following as the reaction temperature-control apparatus
204A, is assigned to the reaction cavity 109 or to a plurality of
reaction cavities 109, in particular in order for it to be possible
to carry out one or more amplification reactions and/or PCRs
therein.
[0131] The reaction cavities 109 are preferably
temperature-controlled simultaneously and/or uniformly, in
particular by means of one common reaction temperature-control
apparatus 204A or two reaction temperature-control apparatuses
204A.
[0132] More particularly preferably, the reaction cavity/cavities
109 can be temperature-controlled from two different sides and/or
by means of two or the reaction temperature-control apparatuses
204A that are preferably arranged on opposite sides.
[0133] Alternatively, for reaction cavities 109, each reaction
cavity 109 can be temperature-controlled independently and/or
individually.
[0134] The temperature-control apparatus 204B, referred to in the
following as the intermediate temperature-control apparatus 204B,
is preferably assigned to the intermediate temperature-control
cavity 110 and/or is designed to temperature-control the
intermediate temperature-control cavity 110 or a fluid located
therein, in particular the amplification products V, preferably to
a preheat temperature TV.
[0135] The intermediate temperature-control cavity 110 and/or
temperature-control apparatus 204B is preferably arranged upstream
of or (immediately) before the sensor apparatus 113, in particular
in order for it to be possible to temperature-control or preheat,
in a desired manner, fluids to be fed to the sensor apparatus 113,
in particular analytes A and/or amplification products V,
particularly preferably immediately before said fluids are fed.
[0136] Particularly preferably, the intermediate
temperature-control cavity 110 and/or temperature-control apparatus
204B is designed or intended to denature the sample P or analytes A
and/or the amplification products V produced, and/or to divide any
double-stranded analytes A or amplification products V into single
strands and/or to counteract premature bonding and/or hybridizing
of the amplification products V, in particular by the addition of
heat.
[0137] The intermediate temperature-control cavity 110 is
preferably elongate and/or designed as a channel which is in
particular sinuous or meandering and/or planar in cross section.
Advantageously, a sufficiently long retention time of the fluid
and/or sufficiently great thermal coupling with the fluid in the
intermediate temperature-control cavity 110 is thus obtained in
order to achieve the desired temperature control for example
without changing the flow speed or also while the fluid is flowing
through said cavity. However, other solutions are also possible
here, in particular those in which the fluid flow in the
intermediate temperature-control cavity 110 is stopped.
[0138] Preferably, the length of the intermediate
temperature-control cavity 110 is at least 10 mm or 15 mm,
particularly preferably at least 20 mm or 25 mm, in particular 30
mm or 40 mm, and/or at most 80 mm or 75 mm, particularly preferably
at most 70 mm or 65 mm, in particular at most 60 mm.
[0139] Preferably, the intermediate temperature-control cavity 110
has a volume of at least 10 .mu.l or 20 .mu.l, particularly
preferably at least 25 .mu.l or 30 .mu.l, and/or at most 500 .mu.l
or 400 .mu.l, particularly preferably at most 350 .mu.l or 300
.mu.l.
[0140] The intermediate temperature-control cavity 110 comprises an
inlet 110A and an outlet 110B, a valve 115, in particular an
initially open valve 115A, preferably being assigned to (each of)
the inlet 110A and/or the outlet 110B, as shown in FIG. 1. In this
way, the flow of fluid through the intermediate temperature-control
cavity 110 can be controlled. For example, it is thus possible to
temperature-control a fluid flowing through the intermediate
temperature-control cavity 110 while it is flowing through and/or
to initially fill the intermediate temperature-control cavity 110
with a fluid to be temperature-controlled and to close the
input-side and/or output-side valve 115A in order to stop the fluid
in the intermediate temperature-control cavity 110 for the purpose
of temperature control and to only subsequently pass on said
fluid.
[0141] Preferably, the intermediate temperature-control cavity 110
is (fluidically) arranged between the reaction cavity/cavities 109
and the sensor apparatus 113 and/or (all) the reaction cavities 109
are fluidically connected or connectable to the sensor apparatus
113, preferably exclusively, by means of the intermediate
temperature-control cavity 110.
[0142] Preferably, the intermediate temperature-control cavity 110
is arranged closer to the sensor apparatus 113 than to the reaction
cavity/cavities 109. In particular, the distance or flow path
between the intermediate temperature-control cavity 110, in
particular the outlet 110B thereof, and the sensor apparatus 113 is
shorter than the distance or flow path between the intermediate
temperature-control cavity 110, in particular the inlet 110A
thereof, and the reaction cavity/cavities 109.
[0143] The intermediate temperature-control cavity 110 is
preferably designed to actively temperature-control, particularly
preferably to heat, fluids, in particular the amplification
products V, preferably to a melting point or melting temperature,
as explained in greater detail in the following.
[0144] The intermediate temperature-control apparatus 204B assigned
to the intermediate temperature-control cavity 110 is preferably
designed to (actively) temperature control, in particular heat, the
intermediate temperature-control cavity 110.
[0145] Preferably, the intermediate temperature-control apparatus
204B comprises a heating element, in particular a heating resistor
or a Peltier element, or is formed thereby.
[0146] The intermediate temperature-control apparatus 204B is
preferably planar and/or has a contact surface which is preferably
elongate and/or rectangular allowing for heat transfer between the
intermediate temperature-control apparatus 204B and the
intermediate temperature-control cavity 110.
[0147] Preferably, the intermediate temperature-control apparatus
204B can be externally positioned against, in particular pressed
against, the cartridge 100, the main body 101 and/or the cover 102,
in the region of the intermediate temperature-control cavity 110 or
on the intermediate temperature-control cavity 110, preferably over
the entire surface thereof.
[0148] In particular, the analysis device 200 comprises the
intermediate temperature-control apparatus 204B. However, other
structural solutions are also possible in which the intermediate
temperature-control apparatus 204B is arranged in the cartridge 100
or integrated in the cartridge 100, in particular in the
intermediate temperature-control cavity 110.
[0149] Preferably, the analysis system 1, analysis device 200
and/or the cartridge 100 and/or one or each temperature-control
apparatus 204 comprise/comprises a temperature detector and/or
temperature sensor (not shown), in particular in order to make it
possible to control and/or regulate temperature.
[0150] One or more temperature sensors may for example be assigned
to the sensor portions 116 and/or to individual channel portions or
cavities, i.e., may be thermally coupled thereto.
[0151] Particularly preferably, a temperature sensor is assigned to
each temperature-control apparatus 204A, 204B and/or 204C, for
example, in order to measure the temperature of the respective
temperature-control apparatuses 204 and/or the contact surfaces
thereof.
[0152] The temperature-control apparatus 204C, referred to in the
following as the sensor temperature-control apparatus 204C, is in
particular assigned to the sensor apparatus 113 and/or is designed
to temperature-control fluids located in or on the sensor apparatus
113, in particular analytes A and/or amplification products V,
reagents or the like, in a desired manner, preferably to a
hybridization temperature TH.
[0153] The sensor temperature-control apparatus 204C preferably
comprises a heating element, in particular a heating resistor or a
Peltier element, or is formed thereby.
[0154] The sensor temperature-control apparatus 204C is preferably
planar and/or has a contact surface which is preferably rectangular
and/or corresponds to the dimensions of the sensor apparatus 113,
the contact surface allowing for heat transfer between the sensor
temperature-control apparatus 204C and the sensor apparatus
113.
[0155] Preferably, the analysis device 200 comprises the sensor
temperature-control apparatus 204C. However, other structural
solutions are also possible in which the sensor temperature-control
apparatus 204C is integrated in the cartridge 100, in particular in
the sensor apparatus 113.
[0156] Particularly preferably, the connection apparatus 203
comprises the sensor temperature-control apparatus 204C, and/or the
connection apparatus 203 together with the sensor
temperature-control apparatus 204C can be linked to, in particular
pressed against, the cartridge 100, in particular the sensor
apparatus 113.
[0157] More particularly preferably, the connection apparatus 203
and the sensor temperature-control apparatus 204C (together) can be
moved toward and/or relative to the cartridge 100, in particular
the sensor apparatus 113, and/or can be positioned against said
cartridge, preferably in order to both electrically and thermally
couple the analysis device 200 to the cartridge 100, in particular
the sensor apparatus 113 or the support 113D thereof.
[0158] Preferably, the sensor temperature-control apparatus 204C is
arranged centrally on the connection apparatus 203 or a support
thereof and/or is arranged between the contact elements 203A.
[0159] In particular, the contact elements 203A are arranged in an
edge region of the connection apparatus 203 or a support thereof or
are arranged around the sensor temperature-control apparatus 204C,
preferably such that the connection apparatus 203 is connected or
connectable to the sensor apparatus 113 thermally in the centre and
electrically on the outside or in the edge region. However, other
solutions are also possible here.
[0160] The analysis system 1 or analysis device 200 preferably
comprises one or more actuators 205 for actuating the valves 115.
Particularly preferably, different (types or groups of) actuators
205A and 205B are provided which are assigned to the different
(types or groups of) valves 115A and 115B for actuating each of
said valves, respectively.
[0161] The analysis system 1 or analysis device 200 preferably
comprises one or more sensors 206. In particular, the sensors 206A
are designed or intended to detect liquid fronts and/or flows of
fluid in the fluid system 103. Particularly preferably, the sensors
206A are designed to measure or detect, for example, optically
and/or capacitively, a liquid front and/or the presence, the speed,
the mass flow rate/volume flow rate, the temperature and/or another
value of a fluid in a channel and/or a cavity, in particular in a
respectively assigned sensor portion 116, which is in particular
formed by a planar and/or widened channel portion of the fluid
system 103.
[0162] Particularly preferably, the sensor portions 116 are each
oriented and/or incorporated in the fluid system 103 and/or fluid
flows against or through the sensor portions 116 such that, in the
operating position of the cartridge 100, fluid flows through the
sensor portions 116 in the vertical direction and/or from the
bottom to the top, in order to make it possible or easier to
reliably detect liquid.
[0163] Alternatively, or additionally, the analysis device 200
preferably comprises (other or additional) sensors 206B for
detecting the ambient temperature, internal temperature,
atmospheric humidity, position, and/or alignment, for example by
means of a GPS sensor, and/or the orientation and/or inclination of
the analysis device 200 and/or the cartridge 100.
[0164] The analysis system 1 or analysis device 200 preferably
comprises a control apparatus 207, in particular comprising an
internal clock or time base for controlling the sequence of a test
and/or for collecting, evaluating and/or outputting or providing
measured values in particular from the sensor apparatus 113, and/or
from test results and/or other data or values.
[0165] The control apparatus 207 preferably controls or regulates
the pump drive 202, the temperature-control apparatuses 204 and/or
actuators 205, in particular taking into account or depending on
the desired test and/or measured values from the sensor apparatus
113 and/or sensors 206.
[0166] Generally, it is noted that the cartridge 100, the fluid
system 103 and/or the conveying of fluid preferably do not operate
on the basis of capillary forces, but at least essentially or
primarily under the effects of gravity and/or the effect of the
pump or pump apparatus 112.
[0167] In the operating position, the liquids from the respective
cavities are preferably removed, in particular drawn out, via the
outlet that is at the bottom in each case, it being possible for
gas or air to flow and/or be pumped into the respective cavities
via the inlet that is in particular at the top. In particular,
relevant vacuums in the cavities can thus be prevented or at least
minimised when conveying the liquids.
[0168] The flows of fluid are controlled in particular by
accordingly activating the pump or pump apparatus 112 and actuating
the valves 115.
[0169] Particularly preferably, the pump drive 202 comprises a
stepper motor, or a drive calibrated in another way, such that
desired metering can be achieved, at least in principle, by means
of appropriate activation.
[0170] Additionally, or alternatively, sensors 206A are preferably
used to detect liquid fronts or flows of fluid, in particular in
cooperation with the assigned sensor portions 116, in order to
achieve the desired fluidic sequence and the desired metering by
accordingly controlling the pump or pump apparatus 112 and
accordingly activating the valves 115.
[0171] Optionally, the analysis system 1 or analysis device 200
comprises an input apparatus 208, such as a keyboard, a touch
screen or the like, and/or a display apparatus 209, such as a
screen.
[0172] The analysis system 1 or analysis device 200 preferably
comprises at least one interface 210, for example for controlling,
for communicating and/or for outputting measured data or test
results and/or for linking to other devices, such as a printer, an
external power supply or the like. This may in particular be a
wired or wireless interface 210.
[0173] The analysis system 1 or analysis device 200 preferably
comprises a power supply 211, preferably a battery or an
accumulator, which is in particular integrated and/or externally
connected or connectable.
[0174] Preferably, an integrated accumulator is provided as a power
supply 211 and is (re)charged by an external charging device (not
shown) via a connection 211A and/or is interchangeable.
[0175] The analysis system 1 or analysis device 200 preferably
comprises a housing 212, all the components and/or some or all of
the apparatuses preferably being integrated in the housing 212.
Particularly preferably, the cartridge 100 can be inserted or slid
into the housing 212, and/or can be received by the analysis device
200, through an opening 213 which can in particular be closed, such
as a slot or the like.
[0176] The analysis system 1 or analysis device 200 is preferably
portable or mobile. Particularly preferably, the analysis device
200 weighs less than 25 kg or 20 kg, particularly preferably less
than 15 kg or 10 kg, in particular less than 9 kg or 6 kg.
[0177] In the following, further details are given on a preferred
construction of the sensor apparatus 113 with reference to FIG. 3
to FIG. 6.
[0178] The sensor apparatus 113 preferably allows electrochemical
measurement and/or redox cycling.
[0179] In particular, the sensor apparatus 113 is designed to
identify, to detect and/or to determine (identical or different)
analytes A bonded to capture molecules M or products derived
therefrom, in particular amplification products V of the analyte A
or different analytes A.
[0180] The sensor apparatus 113 preferably comprises a sensor array
113A comprising a plurality of sensor regions or sensor fields
113B, as shown schematically in FIG. 3, which schematically shows
the measuring side of the sensor apparatus 113 and/or the sensor
array 113A. FIG. 4 is an enlarged detail from FIG. 3. FIG. 5 shows
a connection side and FIG. 6 is a schematic section through the
sensor apparatus 113.
[0181] Preferably, the sensor apparatus 113 or the sensor array
113A comprises more than 10 or 20, particularly preferably more
than 50 or 80, in particular more than 100 or 120 and/or less than
1000 or 800 sensor fields 113B.
[0182] Preferably, the sensor apparatus 113 or the sensor array
113A comprises a plurality of electrodes 113C. At least two
electrodes 113C are preferably arranged in each sensor region or
sensor field 113B. In particular, at least two electrodes 113C in
each case form a sensor field 113B.
[0183] The electrodes 113C are preferably made of metal, in
particular of noble metal, such as platinum or gold, and/or said
electrodes are coated, in particular with thiols.
[0184] Preferably, the electrodes 113C are finger-like and/or
engage in one another, as can be seen from the enlarged detail of a
sensor field 113B according to FIG. 4. However, other structural
solutions or arrangements are also possible.
[0185] The sensor apparatus 113 preferably comprises a support
113D, in particular a chip, the electrodes 113C preferably being
arranged on the support 113D and/or being integrated in the support
113D.
[0186] The measuring side comprises the electrodes 113C and/or is
the side that faces the fluid, the sample P, the amplification
products V and/or a sensor compartment, and/or is the side of the
sensor apparatus 113 and/or the support 113D comprising capture
molecules M (as shown in FIG. 6) to which the analytes A and/or
amplification products V are bonded.
[0187] The connection side of the sensor apparatus 113 and/or the
support 113D is preferably opposite the measuring side and/or is
the side that faces away from the fluid, the sample P and/or the
amplification product V.
[0188] Particularly preferably, the measuring side and the
connection side of the sensor apparatus 113 and/or the support 113D
each form one flat side of the in particular planar and/or
plate-like support 113D.
[0189] The sensor apparatus 113, in particular the support 113D,
preferably comprises a plurality of, in this case eight, electrical
contacts or contact surfaces 113E, the contacts 113E preferably
being arranged on the connection side and/or forming the connection
side, as shown in FIG. 5.
[0190] Preferably, the sensor apparatus 113 can be contacted on the
connection side and/or by means of the contacts 113E and/or can be
electrically connected to the analysis device 200. In particular,
an electrical connection can be established between the cartridge
100, in particular the sensor apparatus 113, and the analysis
device 200, in particular the control apparatus 207, by
electrically connecting the contacts 113E to the contact elements
203A.
[0191] Preferably, the contacts 113E are arranged laterally, in the
edge region and/or in a plan view or projection around the
electrodes 113C and/or the sensor array 113A, and/or the contacts
113E extend as far as the edge region of the sensor apparatus 113,
in particular such that the support 113D can be electrically
contacted, preferably by means of the connection apparatus 203 or
the contact elements 203A, as already explained, laterally, in the
edge region and/or around the sensor temperature-control apparatus
204C, which can preferably be positioned centrally or in the middle
on the support 113D.
[0192] Preferably, the sensor fields 113B are separated from one
another, as shown in the schematic view from FIG. 6. In particular,
the sensor apparatus 113 comprises barriers or partitions between
each of the sensor fields 113B, which are preferably formed by an
in particular hydrophobic layer 113F having corresponding recesses
for the sensor fields 113B. However, other structural solutions are
also possible.
[0193] The cartridge 100 and/or the sensor apparatus 113 comprises
or forms a sensor compartment 113G. In particular, the sensor
compartment 113G is formed between the sensor array 113A, the
sensor apparatus 113 and/or the support 113D, or between the
measuring side on one side and a sensor cover 113H on the other
side.
[0194] The sensor apparatus 113 preferably defines the sensor
compartment 113G by means of its measuring side and/or the sensor
array 113A. The electrodes 113C are therefore in the sensor
compartment 113G.
[0195] Preferably, the cartridge 100 and/or the sensor apparatus
113 comprises the sensor cover 113H, the sensor compartment 113G in
particular being defined or delimited by the sensor cover 113H on
the flat side.
[0196] Particularly preferably, the sensor cover 113H can be
lowered onto the partitions and/or layer 113F for the actual
measurement.
[0197] The sensor apparatus 113 or the sensor compartment 113G is
fluidically linked to the fluid system 103, in particular to the
reaction cavity/cavities 109, preferably by connections 113J, such
that the (treated) sample P, the analytes A or amplification
products V can be admitted to the measuring side of the sensor
apparatus 113 or sensor array 113A.
[0198] The sensor compartment 113G can thus be loaded with fluids
and/or said fluids can flow therethrough.
[0199] The sensor apparatus 113 preferably comprises a plurality of
in particular different capture molecules M, different capture
molecules M preferably being arranged and/or immobilised in or on
different sensor fields 113B and/or preferably being assigned to
different sensor fields 113B.
[0200] Particularly preferably, the electrodes 113C are provided
with capture molecules M, in this case via bonds B, in particular
thiol bonds, in particular in order to bond and/or detect or
identify suitable analytes A and/or amplification products V.
[0201] Different capture molecules M1 to M3 are preferably provided
for the different sensor fields 113B and/or the different electrode
pairs and/or electrodes 113C, in order to specifically bond
different analytes A and/or amplification products V, in FIG. 6 the
amplification products V1 to V3, in the sensor fields 113B.
[0202] Particularly preferably, the sensor apparatus 113 or sensor
array 113A allows the amplification products V bonded in each
sensor field 113B to be qualitatively or quantitatively
determined.
[0203] Preferably, the sensor apparatus 113 comprises capture
molecules M having different hybridization temperatures TH,
preferably in order to bond the amplification products V to the
corresponding capture molecules M at different hybridization
temperatures TH.
[0204] In order to achieve hybridization at the different
hybridization temperatures TH, the temperature of the sensor
apparatus 113, in particular of the electrodes 113C, the support
113D, the sensor compartment 113G and/or the cover 113H, can be
controlled or set, at least indirectly, preferably by means of the
analysis device 200, in particular the sensor temperature-control
apparatus 204B and/or 204C, as already explained.
[0205] Preferably, the sensor temperature-control apparatus 204C is
used to temperature-control the sensor compartment 113G, in this
case by being in contact with the connection side, in particular
such that the desired or required hybridization temperature TH is
reached on the measuring side, in the sensor compartment 113G
and/or in the fluid.
[0206] Preferably, in the operating state, the sensor
temperature-control apparatus 204C rests on the support 113D in a
planar manner and/or centrally and/or so as to be opposite the
sensor array 113A and/or rests on one or more contacts 113E at
least in part. This makes it possible to particularly rapidly and
efficiently temperature-control the sensor compartment 113G and/or
amplification products V.
[0207] The sensor apparatus 113, in particular the support 113D,
preferably comprises at least one, preferably a plurality of,
electronic or integrated circuits, the circuits in particular being
designed to detect electrical currents or voltages that are
preferably generated at the sensor fields 113B in accordance with
the redox cycling principle.
[0208] Particularly preferably, the measurement signals from the
different sensor fields 113B are separately collected or measured
by the sensor apparatus 113 and/or the circuits.
[0209] Particularly preferably, the sensor apparatus 113 and/or the
integrated circuits directly convert the measurement signals into
digital signals or data, which can in particular be read out by the
analysis device 200.
[0210] Particularly preferably, the sensor apparatus 113 and/or the
support 113D is constructed as described in European Patent EP 1
636 599 B1 and corresponding U.S. Pat. No. 7,914,655.
[0211] In the following, a preferred sequence of a test or analysis
using the proposed analysis system 1 and/or analysis device 200
and/or the proposed cartridge 100 and/or in accordance with the
proposed method is explained in greater detail by way of
example.
[0212] The analysis system 1, the cartridge 100 and/or the analysis
device 200 is preferably designed to carry out the proposed
method.
[0213] During the proposed method for testing a sample P, at least
one analyte A of the sample P is preferably amplified or copied, in
particular by means of PCR. The amplified analyte A and/or the
amplification products V produced in this way is/are then bonded
and/or hybridized to corresponding capture molecules M. The bonded
amplification products V are then detected, in particular by means
of electronic measurement.
[0214] The method may be used in particular in the field of
medicine, in particular veterinary medicine, in order to detect
diseases and/or pathogens.
[0215] Within the context of the method according to the invention,
a sample P having at least one analyte A on the basis of a fluid or
a liquid from the human or animal body, in particular blood, saliva
or urine, is usually first introduced into the receiving cavity 104
via the connection 104A, in order to detect diseases and/or
pathogens, it being possible for the sample P to be
pre-treated.
[0216] Once the sample P has been received, the receiving cavity
104 and/or the connection 104A thereof is fluidically closed, in
particular in a liquid-tight and/or gas-tight manner.
[0217] Preferably, the cartridge 100 together with the sample P is
then linked or connected to the analysis device 200, in particular
is inserted or slid into the analysis device 200.
[0218] The method sequence, in particular the flow and conveying of
the fluids, the mixing and the like, is controlled by the analysis
device 200 or the control apparatus 207, in particular by
accordingly activating and actuating the pump drive 202 or the pump
apparatus 112 and/or the actuators 205 or valves 115.
[0219] Preferably, the sample P, or a part or supernatant of the
sample P, is removed from the receiving cavity 104 via the outlet
104C and/or the intermediate connection 104D and is fed to the
mixing cavity 107 in a metered manner.
[0220] Preferably, the sample P in the cartridge 100 is metered, in
particular in or by means of the first metering cavity 105A and/or
second metering cavity 105B, before being introduced into the
mixing cavity 107. Here, in particular the upstream and/or
downstream sensor portions 116 are used together with the assigned
sensors 206 in order to make possible the desired metering.
However, other solutions are also possible.
[0221] In the mixing cavity 107, the sample P is prepared for
further analysis and/or is mixed with a reagent, preferably with a
liquid reagent F1 from a first storage cavity 108A and/or with one
or more dry reagents S1, S2 and/or S3, which are preferably
provided in the mixing cavity 107.
[0222] The liquid and/or dry reagents can be introduced into the
mixing cavity 107 before and/or after the sample P. In the example
shown, the dry reagents S1 to S3 are preferably introduced into the
mixing cavity 107 previously and are optionally dissolved by the
sample P and/or the liquid reagent F1.
[0223] The liquid reagent F1 may in particular be a reagent, in
particular a PCR master mix, for the amplification reaction or PCR.
Preferably, the PCR master mix contains nuclease-free water,
enzymes for carrying out the PCR, in particular at least one DNA
polymerase, nucleoside triphosphates (NTPs), in particular
deoxynucleotides (dNTPs), salts, in particular magnesium chloride,
and/or reaction buffers.
[0224] The dry reagents S1, S2 and/or S3 may likewise be reagents
required for carrying out an amplification reaction or PCR, which
are in a dry, in particular lyophilised, form. Preferably, the dry
reagents S1, S2 and/or S3 are selected in particular from
lyophilised enzymes, preferably DNA polymerases, NTPs, dNTPs and/or
salts, preferably magnesium chloride.
[0225] The dissolving or mixing in the mixing cavity 107 takes
place or is assisted in particular by introducing and/or blowing in
gas or air, in particular from the bottom. This is carried out in
particular by accordingly pumping gas or air in the circuit by
means of the pump or pump apparatus 112.
[0226] Subsequently, a desired volume of the sample P that is mixed
and/or pretreated in the mixing cavity 107 is preferably fed to one
or more reaction cavities 109, particularly preferably via
(respectively) one of the upstream, optional intermediate cavities
106A to 106C and/or with different reagents or primers, in this
case dry reagents S4 to S6, being added or dissolved.
[0227] Particularly preferably, the (premixed) sample P is split
into several sample portions, preferably of equal size, and/or is
divided between the intermediate cavities 106A to 106C and/or
reaction cavities 109, preferably evenly and/or in sample portions
of equal size.
[0228] Different reagents, in the present case dry reagents S4 to
S6, particularly preferably primers, in particular those required
for the PCR or PCRs, in particular groups of different primers in
this case, are preferably added to the (premixed) sample P in the
intermediate cavities 106A to 106C and/or different reaction
cavities 109, respectively.
[0229] The primers in the different groups differ in particular in
terms of the hybridization temperatures of the amplification
products V produced by the respective primers. As a result, in
particular the different group temperatures of the groups of
analytes A and/or amplification products V are produced, as already
mentioned at the outset.
[0230] Particularly preferably, marker primers are used in the
sense already specified at the outset.
[0231] In the embodiment shown, the reagents or primers S4 to S6
are contained in the intermediate cavities 106A to 106C. However,
other solutions are also possible, in particular those in which the
reagents or primers S4 to S6 are contained in the reaction cavities
109.
[0232] According to a preferred embodiment, the intermediate
cavities 106A to 106C each contain primers for amplifying/copying
one analyte A, preferably two different analytes A and more
preferably three different analytes A. However, it is also possible
for four or more different analytes A to be amplified/copied per
reaction cavity 109.
[0233] Particularly preferably, the reaction cavities 109 are
filled in succession with a specified volume of the (pre-treated)
sample P or with respective sample portions via the intermediate
cavities 106A to 106C that are each arranged upstream. For example,
the first reaction cavity 109A is filled with a specified volume of
the pre-treated sample P before the second reaction cavity 109B
and/or the second reaction cavity 109B is filled therewith before
the third reaction cavity 109C.
[0234] In the reaction cavities 109, the amplification reactions or
PCRs are carried out to copy/amplify the analytes A. This is
carried out in particular by means of the assigned, preferably
common, reaction temperature-control apparatus 204A and/or
preferably simultaneously for all the reaction cavities 109, i.e.,
in particular using the same cycles and/or temperature
(curves/profiles).
[0235] The PCR or PCRs are carried out on the basis of protocols or
temperature profiles that are essentially known to a person skilled
in the art. In particular, the mixture or sample volume located in
the reaction cavities 109 is preferably cyclically heated and
cooled.
[0236] Preferably, nucleic-acid products are produced from the
analytes A as amplification products V in the reaction
cavity/cavities 109.
[0237] During the pretreatment, reaction and/or PCR or
amplification, a label L is directly produced (in each case) and/or
is attached to the amplification products V. This is in particular
achieved by using corresponding, preferably biotinylated, primers.
However, the label L can also be produced and/or bonded to the
amplification products V separately or later, optionally also only
in the sensor compartment 113G and/or after hybridization.
[0238] The label L is used in particular for detecting bonded
amplification products V. In particular, the label L can be
detected or the label L can be identified in a detection process,
as explained in greater detail in the following.
[0239] According to the invention, it is possible for a plurality
of amplification reactions or PCRs to be carried out in parallel
and/or independently from one another using different primers S4 to
S6 and/or primer pairs, such that a large number of (different)
analytes A can be copied or amplified in parallel and subsequently
analysed.
[0240] In particular, identical or different analytes A1 are
amplified in the first reaction cavity 109A, identical or different
analytes A2 are amplified in the second reaction cavity 109B and
identical or different analytes A3 are amplified in the third
reaction cavity 109C, preferably by means of amplification
reactions, in particular PCRs, that run in parallel.
[0241] Particularly preferably, the analytes A1 to A3 are different
from one another, in particular such that a large number of
different analytes A can be amplified and/or tested by means of the
method. Preferably, more than 2 or 4, particularly preferably more
than 8 or 11, in particular more than 14 or 17, analytes A can be
tested and/or amplified, in particular at the same time.
[0242] In particular, a plurality of groups of amplification
products V of the analytes A are formed and/or produced, preferably
in parallel and/or independently from one another and/or in the
reaction cavities 109. Therefore, for example, a first group of
amplification products V1 of the analytes A1 is formed and/or
produced in the first reaction cavity 109A, a second group of
amplification products V2 of the analytes A2 is formed and/or
produced in the second reaction cavity 109B, and a third group of
amplification products V3 of the analytes A3 is formed and/or
produced in the optional third reaction cavity 109C.
[0243] Particularly preferably, groups of (amplified) analytes A
and/or amplification products V are formed that have different
group temperatures in the sense mentioned at the outset. The groups
thus preferably have different (optimal) hybridization temperatures
TH and/or ranges of hybridization temperatures.
[0244] Preferably, different groups of analytes A and/or
amplification products V, i.e., in particular nucleic-acid products
and/or sequences, are thus amplified and/or formed for the test, it
being possible, for the different groups to be amplified and/or
formed and/or provided in particular in the different reaction
chambers 109A to 109C, but alternatively also in a different
manner.
[0245] After carrying out the PCR and/or amplification,
corresponding fluid volumes and/or amplification products V and/or
the groups are conducted out of the reaction cavities 109 in
succession to the sensor apparatus 113 and/or to the sensor
compartment 113G, in particular via a group-specific and/or
separate intermediate cavity 106E, 106F or 106G (respectively)
and/or via the optional (common) intermediate temperature-control
cavity 110.
[0246] The intermediate cavities 106E to 106G may contain further
reagents, in this case dry reagents S9 and S10, respectively, for
preparing the amplification products V for the hybridization, e.g.
a buffer, in particular an SSC buffer, and/or salts for further
conditioning. On this basis, further conditioning of the
amplification products V can be carried out, in particular in order
to improve the efficiency of the subsequent hybridization (bonding
to the capture molecules M). Particularly preferably, the pH of the
sample P is set or optimised in the intermediate cavities 106E to
106G and/or by means of the dry reagents S9 and S10.
[0247] Preferably, the sample P or the analytes A and/or
amplification products V or groups formed thereby is/are, in
particular immediately before being fed to the sensor apparatus 113
and/or between the reaction cavities 109 and the sensor apparatus
113, actively temperature-controlled (in particular in advance
and/or before being temperature-controlled in the sensor apparatus
113), preferably preheated, in particular by means of and/or in the
intermediate temperature-control cavity 110 and/or by means of the
intermediate temperature-control apparatus 204B.
[0248] Preferably, the groups and/or analytes A or amplification
products V of the individual reaction cavities 109 are actively
temperature-controlled (in particular in advance and/or before
being temperature-controlled in the sensor apparatus 113) and/or
fed to the intermediate temperature-control cavity 110 in
succession. The groups are in particular fed to the sensor
apparatus 113 and/or the sensor compartment 113G in succession
being temperature-controlled, in particular in advance and/or
before being temperature-controlled in the sensor apparatus
113.
[0249] FIG. 7 shows an exemplary schematic curve or profile for the
temperature T of the sample P as a function of the position X in or
on the cartridge 100.
[0250] The sample P is preferably fed to the cartridge 100 and/or
receiving cavity 104 at or with an ambient temperature TU, for
example of approximately 20.degree. C. The amplification reactions
are then carried out in the reaction cavities 109, the prepared
sample P preferably being cyclically heated and cooled (not shown
in FIG. 7).
[0251] As already explained, a plurality of groups having in
particular different analytes A and/or amplification products V
and/or group temperatures or hybridization temperatures TH are
preferably produced.
[0252] The groups and/or amplification products V are then
preferably fed to the assigned intermediate cavities 106E to 106G
and/or to the subsequent intermediate temperature-control cavity
110, preferably in succession.
[0253] Preferably, the groups and/or amplification products V cool
at different rates and/or continuously in the reaction cavities
109, and/or the groups and/or amplification products V leave the
reaction cavities 109 in succession and/or at different
temperatures, as shown schematically in FIG. 7. However, other
method variants are also possible in which the groups and/or
amplification products V are also temperature-controlled and/or
kept at a constant temperature in the reaction cavities 109 after
the end of the PCRs, preferably such that the groups and/or
amplification products V leave the reaction cavities 109 at the
same temperature.
[0254] Preferably, the groups and/or amplification products V cool
on the way to the intermediate temperature-control cavity 110. In
this process, the groups and/or amplification products V can cool
particularly significantly and/or additionally in the intermediate
cavities 106B to 106G by absorbing the reagents S9 and S10
contained in said cavities, as shown in FIG. 7 by a jump in the
temperature curve or temperature profile between the reaction
cavities 109 and the inlet 110A of the intermediate
temperature-control cavity 110, and/or at the reaction cavities
106.
[0255] Preferably, the groups and/or amplification products V have
different inlet temperatures TE at the inlet 110A of the
intermediate temperature-control cavity 110, as shown in FIG. 7 at
the inlet 110A, in particular if they have left the reaction
cavities 109A to 109C at different temperatures. However, the inlet
temperatures TE may also be substantially identical.
[0256] The inlet temperature TE at the inlet 110A preferably
corresponds at least substantially to the ambient temperature TU or
is at most 10.degree. C. or 5.degree. C. above the ambient
temperature TU. However, the inlet temperature TE may also be
higher if necessary.
[0257] Preferably, the groups and/or amplification products V are
heated (in succession) in the intermediate temperature-control
cavity 110 to a preheat temperature TV and/or melting point or
melting temperature, the preheat temperature TV preferably being
reached (at the latest) at the outlet 110B of the intermediate
temperature-control cavity 110.
[0258] Preferably, the preheat temperature TV is higher than the
hybridization temperature TH, and in particular at least as high as
the melting point or melting temperature of the respective groups
and/or amplification products V. In particular, the groups and/or
amplification products V are heated to the preheat temperature TV
immediately before being fed to the sensor apparatus 113 and/or
between the reaction cavities 109 and the sensor apparatus 113, in
particular in order to denature the groups and/or amplification
products V, as already explained.
[0259] As shown in FIG. 7, all the groups and/or amplification
products V are preferably heated to the same preheat temperature
TV, for example at least 95.degree. C.
[0260] However, other method variants are also possible in which
the groups and/or amplification products V are
temperature-controlled (in particular in advance and/or before
being temperature-controlled in the sensor apparatus 113) and/or
(pre-)heated to different preheat temperatures TV. In particular,
the preheat temperature TV can be varied for each group and/or
depending on the required hybridization temperature TH and/or group
temperature. In particular, the preheat temperature TV of the first
group may be greater than the preheat temperature TV of the second
and/or third group and/or the preheat temperature TV may decrease
from group to group.
[0261] The melting point or melting temperature and/or preheat
temperature TV is preferably above the respective hybridization
temperatures TH and/or is at least 70.degree. C. or 80.degree. C.
and/or at most 99.degree. C. or 96.degree. C., in particular such
that bonds of the analytes A and/or amplification products V
produced in the meantime dissolve, and/or such that the analytes A
and/or amplification products V can be fed to the sensor apparatus
113 in the denatured and/or dissolved state.
[0262] Optionally, the analytes A and/or amplification products V
and/or the groups of amplification products V are
temperature-controlled (in particular in advance and/or before
being temperature-controlled in the sensor apparatus 113), in
particular (pre-) heated, to the corresponding hybridization
temperature TH before being fed to the sensor apparatus 113,
preferably such that they can be bonded directly to the
corresponding capture molecules M after being fed to the sensor
apparatus 113.
[0263] In an alternative method variant, the groups and/or
amplification products V are actively temperature-controlled, in
particular heated, (exclusively) in or on the sensor apparatus 113,
and/or brought to the corresponding hybridization temperature TH,
preferably solely by means of the sensor temperature-control
apparatus 204C. In particular, both the denaturing of any
hybridized amplification products V and the (subsequent)
hybridization of the amplification products V and the corresponding
capture molecules M can take place in or on the sensor apparatus
113. In this case, previous (intermediate) temperature control
before the sensor apparatus 113 can therefore be omitted.
[0264] In the preferred method variant, the sample P and/or the
groups or analytes A and/or amplification products V is/are,
however, in particular immediately before being fed to the sensor
apparatus 113 and/or between the reaction cavities 109 and the
sensor apparatus 113, actively temperature-controlled (in
particular in advance and/or before being temperature-controlled in
the sensor apparatus 113) and/or brought to the preheat temperature
TV, preferably by means of the intermediate temperature-control
apparatus 204B, and, after being fed to the sensor apparatus 113
and/or in the sensor apparatus 113, is/are subsequently and/or
again temperature-controlled (in particular after being
temperature-controlled in the intermediate temperature-control
cavity 110) and/or brought to the corresponding hybridization
temperature TH and/or group temperature, preferably by means of the
sensor temperature-control apparatus 204C. In this case, any
hybridized amplification products V are thus denatured before being
fed to and/or outside the sensor apparatus 113.
[0265] In particular, in the preferred method variant, the sample P
and/or the groups and/or amplification products V is/are brought to
the respective hybridization temperatures TH and/or group
temperatures in multiple stages or more rapidly after leaving the
reaction cavity/cavities 109, preferably the amplification products
V being, in a first stage, temperature-controlled, in particular in
the intermediate temperature-control cavity 110 and/or in advance
and/or before being temperature-controlled in the sensor apparatus
113, to a temperature above the hybridization temperature TH and/or
to the preheat temperature TV and/or being denatured at the melting
point or melting temperature, and, in a second stage, being
subsequently and/or again temperature-controlled, in particular
heated and/or cooled, to the corresponding hybridization
temperature TH and/or group temperature, in particular in the
sensor apparatus 113 and/or after being temperature-controlled in
the intermediate temperature-control cavity 110.
[0266] By means of the sensor temperature-control apparatus 204C,
the sensor apparatus 113 is in particular preheated such that in
particular undesired cooling of the sample P that is preheated, in
this case in the intermediate temperature-control cavity 110,
and/or groups, in particular to below the respective hybridization
temperatures TH and/or group temperatures, can be prevented.
[0267] Particularly preferably, the sensor apparatus 113 is
preheated in each case at least substantially to the hybridization
temperature TH of the respective analytes A and/or amplification
products V, and/or to the respective group temperatures or to a
slightly higher or lower temperature. Owing to the relatively large
thermal mass of the sensor apparatus 113, the desired and/or
optimal temperature for the hybridization can be (rapidly) reached
when the preferably warmer sample P and/or group is fed into the
sensor apparatus 113 and/or the sensor compartment 113G
thereof.
[0268] The amplification products V, nucleic-acid products and/or
the groups from the reaction cavities 109 are conducted to the
sensor apparatus 113 in succession, in particular in order to be
detected or determined therein.
[0269] Preferably, the first group and/or the amplification
products V1 from the first reaction cavity 109A is/are fed to the
sensor apparatus 113 and/or bonded to the corresponding capture
molecules M1 before the second group and/or the amplification
products V2 from the second reaction cavity 109B, in particular the
second group and/or the amplification products V2 from the second
reaction cavity 109B being bonded to the corresponding capture
molecules M2 before the third group and/or the amplification
products V3 from the third reaction cavity 109C.
[0270] After the sample P and/or the amplification products V are
fed to the sensor apparatus 113, the amplification products V are
hybridized to the capture molecules M.
[0271] In the context of the present invention, it has proven to be
particularly advantageous to hybridize the amplification products V
and/or groups of the amplification products V at a hybridization
temperature TH and/or group temperature that is specifically
selected in each case.
[0272] Particularly preferably, the sample portions and/or
amplification products V from the different PCRs and/or from the
different reaction cavities 109 are bonded to the capture molecules
M, in particular in succession, at different hybridization
temperatures TH and/or at decreasing hybridization temperatures TH
and/or group temperatures.
[0273] Preferably, the analytes A and/or amplification products V
in a group each have a similar, preferably at least substantially
identical, (optimal) hybridization temperature TH at which they
bond to the suitable capture molecules M. However, it is also
possible for the analytes A and/or amplification products V in a
group to each have somewhat different (optimal) hybridization
temperatures TH, i.e., a range of hybridization temperatures, as
already explained at the outset. Therefore, this results in an
average and/or optimal hybridization temperature TH of the group or
a temperature range of (optimal) hybridization temperatures for
this group. This hybridization temperature TH of the group or this
temperature range is also referred to as the "group temperature"
for short.
[0274] The groups and/or amplification products V can be hybridized
at a decreasing or increasing, preferably decreasing, group
temperature and/or hybridization temperature TH. If a decreasing
hybridization temperature TH is used, amplification products V that
are already bonded can be prevented from becoming detached from the
capture molecules M again due to the subsequent temperature
increase.
[0275] Particularly preferably, the group temperature and/or
hybridization temperature TH1 of the first group, amplification
products V1 and/or analytes A1 is greater than the group
temperature and/or hybridization temperature TH2 of the second
group, amplification products V2 and/or analytes A2, and this
temperature is in turn greater than the third group temperature
and/or hybridization temperature TH3 of the third group,
amplification products V3 and/or analytes A3.
[0276] "Hybridization temperature" is understood to mean in
particular the temperature at which, on average, the most analytes
A and/or amplification products V in the respective groups bond to
the suitable capture molecules M.
[0277] The group temperatures and/or hybridization temperatures TH
of the different groups preferably differ by more than 3.degree.
C., in particular more than 4.degree. C., preferably by
approximately 5.degree. C. or more.
[0278] Preferably, the group temperature and/or hybridization
temperature TH is at least 40.degree. C. or 45.degree. C. and/or at
most 75.degree. C. or 70.degree. C.
[0279] Preferably, the first group temperature and/or hybridization
temperature TH1 of the first group and/or amplification products V1
is at least 55.degree. C. or 58.degree. C., particularly preferably
at least 60.degree. C. or 62.degree. C., and/or at most 80.degree.
C. or 78.degree. C., particularly preferably at most 75.degree. C.
or 72.degree. C.
[0280] Preferably, the second group temperature and/or
hybridization temperature TH2 of the second group and/or
amplification products V2 is at least 40.degree. C. or 45.degree.
C., particularly preferably at least 48.degree. C. or 52.degree.
C., and/or at most 70.degree. C. or 65.degree. C., particularly
preferably at most 60.degree. C. or 58.degree. C.
[0281] Preferably, the third group temperature and/or hybridization
temperature TH3 of the third group and/or amplification products V3
is at least 35.degree. C. or 40.degree. C., particularly preferably
at least 42.degree. C. or 45.degree. C., and/or at most 65.degree.
C. or 62.degree. C., particularly preferably at most 60.degree. C.
or 55.degree. C.
[0282] At the first group temperature and/or hybridization
temperature TH1, for example approximately 60.degree. C., the first
group bonds particularly well to the corresponding or suitable
capture molecules M1. At the second group temperature and/or
hybridization temperature TH2, for example approximately 55.degree.
C., the second group bonds particularly well to the corresponding
or suitable capture molecules M2. At the third group temperature
and/or hybridization temperature TH3, for example approximately
50.degree. C., the third group bonds particularly well to the
corresponding or suitable capture molecules M3.
[0283] As shown in FIG. 7, the groups and/or amplification products
V cool on the way from the intermediate temperature-control cavity
110 to the sensor apparatus 113. Depending on the temperature
control in advance and/or in the intermediate temperature-control
cavity 110, the preheat temperature TV and/or the temperature
prevailing when the fluid enters the sensor apparatus 113, and/or
depending on the group temperature and/or optimal hybridization
temperature TH, it may therefore be necessary to
temperature-control individual or all groups and/or amplification
products V or the sensor apparatus 113 to different extents by
means of the sensor temperature-control apparatus 204C.
[0284] For example, the first group and/or the amplification
products V1 from the first reaction cavity 109A is/are
temperature-controlled, in particular heated, to a greater extent
or cooled to a lesser extent than the second group and/or the
amplification products V2 from the second reaction cavity 109B
and/or the third group and/or amplification products V3 from the
third reaction cavity 109C.
[0285] In particular, the temperature control of the sensor
apparatus 113, in particular of the support 113D, is adapted for
each group and/or the different amplification products V in order
to reach the respective group temperatures and/or hybridization
temperatures TH.
[0286] In the example shown, the hybridization temperature TH1 of
the first group is above the temperature of the first group at
which it enters the sensor apparatus 113, preferably such that the
first group and/or the amplification products V1 from the first
reaction cavity 109A has/have to be heated for the hybridization,
for example by more than 2.degree. C. or 5.degree. C.
[0287] The hybridization temperature TH may, however, also
correspond to the inlet temperature TE or temperature at entry into
the sensor apparatus 113. In this case, the respective groups
and/or the amplification products V are kept at a constant
temperature in or on the sensor apparatus 113 for the
hybridization. In particular, the group and/or the amplification
products V may already be fed to the sensor apparatus 113 at the
corresponding hybridization temperature TH, as shown in FIG. 7 for
the second group and/or the amplification products V2 from the
second reaction cavity 109B.
[0288] Furthermore, it is possible that the inlet temperature TE or
temperature at entry into the sensor apparatus 113 is greater than
the hybridization temperature TH of the respective groups and/or of
the amplification products V. In this case, the respective groups
and/or the amplification products V are cooled or (slightly)
temperature-controlled in or on the sensor apparatus 113 for the
hybridization such that the temperature is reduced to the required
hybridization temperature TH, in particular at a specified speed,
as shown in FIG. 7 for the third group and/or the amplification
products V3 from the third reaction cavity 109C.
[0289] According to the invention, it may be provided both that the
hybridization temperature TH is changed in stages, for example in
increments of several degrees Celsius and/or in 5.degree. C.
increments, and that the hybridization temperature TH is changed,
in particular reduced, continuously and/or gradually during the
hybridization of a group or at least one analyte A and/or
amplification product V.
[0290] In another method variant, it may be provided that the
respective groups and/or amplification products V in the respective
groups are temperature-controlled differently, and/or the
temperature is varied in or on the sensor apparatus 113 for the
hybridization of the amplification products V in one of the groups,
preferably in order to bond the different amplification products V
in the respective groups to the corresponding capture molecules M
at respectively different hybridization temperatures TH.
[0291] Once the sample P, groups, analytes A and/or amplification
products V are hybridized and/or bonded to the capture molecules M,
detection follows, in particular by means of the preferably
provided labels L, or in another manner.
[0292] In the following, a particularly preferred variant of the
detection is described in greater detail, specifically
electrochemical detection, but other types of detection, for
example optical detection, capacitive detection or the like, may
also be carried out.
[0293] Following the respective bondings/hybridizations, preferably
an optional washing process takes place and/or additional reagents
or liquids, in particular from the storage cavities 108B to 108E,
are optionally fed in.
[0294] In particular, it may be provided that sample residues
and/or unbonded amplification products V, reagents and/or remnants
of the PCR and other substances that may disrupt the rest of the
method sequence are removed.
[0295] Washing or flushing may in particular take place using a
fluid and/or reagent F3, in particular a wash buffer, particularly
preferably a sodium-citrate buffer or SSC buffer, which is
preferably contained in the storage cavity 108C. Unbonded analytes
A and/or amplification products V, and substances which could
disrupt subsequent detection, are preferably removed from the
sensor apparatus 113 and/or fed to the collection cavity 111 by the
wash buffer.
[0296] Subsequently and/or after the washing process, in accordance
with a preferred variant of the method, detection of the
amplification products V bonded to the capture molecules M takes
place.
[0297] In order to detect the amplification products V bonded to
the capture molecules M, a reagent F4 and/or detector molecules D,
in particular alkaline phosphatase/streptavidin, is/are fed to the
sensor apparatus 113, preferably from the storage cavity 108D.
[0298] The reagents F4 and/or detector molecules D can bond to the
bonded amplification products V, in particular to the label L of
the bonded amplification products V, particularly preferably to the
biotin marker, as shown in FIG. 6.
[0299] In the context of detection, it may also be provided that
additional liquid reagents F3 and/or F5 are fed from the storage
cavities 108C and/or 108E to the sensor apparatus 113.
[0300] Optionally, subsequently or after the reagents F4 and/or
detector molecules D have bonded to the amplification products V
and/or the labels L, an (additional) washing process and/or
flushing takes place, preferably by means of the fluid and/or
reagent F3 and/or wash buffer, in particular in order to remove
unbonded reagents F4 and/or detector molecules D from the sensor
apparatus 113.
[0301] Preferably, a reagent S7 and/or S8 and/or substrate SU for
the detection, in particular from the storage cavity 106D, is
lastly fed to the sensor apparatus 113, preferably together with a
fluid or reagent F2 (in particular a buffer), which is suitable for
the substrate SU, particularly preferably for dissolving the
reagent S7 and/or S8 and/or substrate SU, the fluid or reagent F2
in particular being taken from the storage cavity 106B. In
particular, the reagent S7 and/or S8 can form or can comprise the
substrate SU.
[0302] After adding the substrate SU, the cover 113H is preferably
lowered in order to isolate the sensor fields 113B from one another
and/or to minimise the exchange of substances therebetween.
[0303] Preferably, p-aminophenyl phosphate (pAPP) is used as the
substrate SU.
[0304] The substrate SU preferably reacts on and/or with the bonded
amplification products V and/or detector molecules D and/or allows
these to be electrochemically measured.
[0305] Preferably, the substrate SU is split by the bonded detector
molecules D, in particular the alkaline phosphatase of the bonded
detector molecules D, preferably into a first substance SA, such as
p-aminophenol, which is in particular electrochemically active
and/or redox active, and a second substance SP, such as
phosphate.
[0306] Preferably, the first or electrochemically active substance
SA is detected in the sensor apparatus 113 or in the individual
sensor fields 113B by electrochemical measurement and/or redox
cycling.
[0307] Particularly preferably, by means of the first substance SA,
specifically a redox reaction takes place at the electrodes 113C,
the first substance SA preferably discharging electrons to or
receiving electrons from the electrodes 113C.
[0308] In particular, the presence of the first substance SA and/or
the respective amounts in the respective sensor fields 113B is
detected by the associated redox reactions. In this way, it can be
determined qualitatively and in particular also quantitatively
whether and how many analytes A and/or amplification products V are
bonded to the capture molecules M in the respective sensor fields
113B. This accordingly gives information on which analytes A are or
were present in the sample P, and in particular also gives
information on the quantity of said analytes A.
[0309] In particular, by means of the redox reaction with the first
substance SA, an electrical current signal or power signal is
generated at the assigned electrodes 113C, the current signal or
power signal preferably being detected by means of an assigned
electronic circuit.
[0310] Depending on the current signal or power signal from the
electrodes 113C that is generated in this way, it is determined
whether and/or where hybridization to the capture molecules M has
occurred.
[0311] The measurement is preferably taken just once and/or for the
entire sensor array 113A and/or for all the sensor fields 113B, in
particular simultaneously or in parallel. In particular, the bonded
groups and/or amplification products V from all the groups and/or
reaction cavities 109 are detected, identified or determined
simultaneously or in parallel in a single or common detection
process.
[0312] In other words, the amplification products V from the
individual reaction cavities 109 that are bonded at different
and/or specifically selected hybridization temperatures TH are
detected together and/or in parallel, such that rapid measurement
is possible, and high specificity in relation to the hybridization
of the analytes A and/or amplification products V to the capture
molecules M is nevertheless also achieved on the basis of the
hybridization temperature TH that is set in a targeted manner in
each case.
[0313] However, in principle, it is also possible to measure a
plurality of sample portions in the sensor apparatus 113 or in a
plurality of sensor apparatuses 113 in succession or
separately.
[0314] The test results or measurement results are in particular
electrically transmitted to the analysis device 200 or the control
apparatus 207 thereof, preferably by means of the electrical
connection apparatus 203, and are accordingly prepared, analysed,
stored, displayed and/or output, in particular by the display
apparatus 209 and/or interface 210.
[0315] After the test has been carried out, the cartridge 100 is
disconnected from the analysis device 200 and/or is released and/or
ejected therefrom, and is in particular disposed of.
[0316] Individual aspects and features of the present invention and
individual method steps and/or method variants may be implemented
independently from one another, but also in any desired combination
and/or order.
[0317] In particular, the present invention relates also to any one
of the following aspects which can be realized independently or in
any combination, also in combination with any aspects described
above.
1. Analysis system (1) for testing an in particular biological
sample (P), the analysis system (1) comprising a receiving cavity
(104) for the sample (P) and/or a reaction cavity (109) for forming
amplification products (V) of analytes (A) of the sample (P) and
further comprising a sensor apparatus (113) for detecting the
analytes (A) and/or amplification products (V), the sensor
apparatus (113) being fluidically connected to the receiving cavity
(104) and/or reaction cavity (109), characterized in that the
analysis system (1) comprises an intermediate temperature-control
cavity (110) for actively temperature-controlling the analytes (A)
and/or amplification products (V), the intermediate
temperature-control cavity (110) being arranged between the
receiving cavity (104) and/or reaction cavity (109) on one side and
the sensor apparatus (113) on the other side, and/or in that the
sensor apparatus (113) comprises a support (113D) and a plurality
of electrodes (113C) arranged on the support (113D), and the
analysis system (1) comprises a sensor temperature-control
apparatus (204C) for in particular directly temperature-controlling
the support (113D). 2. Analysis system according to aspect 1,
characterised in that the analysis system (1) comprises a plurality
of reaction cavities (109) for producing the amplification products
(V) in parallel and/or independently, and/or in that the sensor
apparatus (113) comprises capture molecules (M) for bonding the
amplification products (V), the sensor apparatus (113) preferably
being fluidically connected to all the reaction cavities (109) via
the intermediate temperature-control cavity (110). 3. Analysis
system according to aspect 1 or 2, characterised in that the
intermediate temperature-control cavity (110) is elongate and/or
designed as a preferably sinuous channel 4. Analysis system
according to any of the preceding aspects, characterised in that
the analysis system (1) comprises an intermediate
temperature-control apparatus (204B) for actively
temperature-controlling the intermediate temperature-control cavity
(110), preferably the intermediate temperature-control apparatus
(204B) comprising a heating resistor or a Peltier element or being
formed thereby. 5. Analysis system according to any one of the
preceding aspects, characterised in that the support (113D) is
arranged between the electrodes (113C) and the sensor
temperature-control apparatus (204C), in that, in the operating
state, the sensor temperature-control apparatus (204C) rests on the
support (113D), preferably in a planar manner and/or centrally,
and/or in that the sensor temperature-control apparatus (204C)
comprises a heating resistor or a Peltier element or is formed
thereby. 6. Analysis system according to any one of the preceding
aspects, characterised in that the support (113D) comprises a chip
or is formed by a chip, the chip preferably being electrically
contactable and/or comprising a plurality of electrical contacts
(113E), in particular laterally, in the edge region and/or around
the sensor temperature-control apparatus (204C). 7. Analysis system
according to any one of the preceding aspects, characterised in
that the analysis system (1) comprises a connection apparatus (203)
for electrically and/or thermally connecting the sensor apparatus
(113), in particular the support (113D), preferably the connection
apparatus (203) comprising the sensor temperature-control apparatus
(204C) and/or a plurality of electrical contact elements (203A)
and/or being able to be moved, in particular pressed, against the
sensor apparatus (113), in particular the support (113D), or vice
versa. 8. Analysis system according to any one of the preceding
aspects, characterised in that the analysis system (1) comprises a
cartridge (100) for receiving the sample (P) and an analysis device
(200) for receiving the cartridge (100), preferably the cartridge
(100) comprising the receiving cavity (104), reaction
cavity/cavities (109), sensor apparatus (113) and/or intermediate
temperature-control cavity (119), and/or the analysis device (200)
comprising the sensor temperature-control apparatus (204C),
intermediate temperature-control apparatus (204B) and/or the
connection apparatus (203). 9. Method for testing an in particular
biological sample (P), analytes (A) of the sample (P) being
pretreated and/or amplification products (V) being produced from
analytes (A) of the sample (P) in a reaction cavity (109), and the
pretreated analytes (A) and/or amplification products (V) being
bonded to capture molecules (M) on a support (113D) of a sensor
apparatus (113) and the bonded analytes (A) and/or amplification
products (V) being detected by means of the sensor apparatus (113),
characterized in that the amplification products (V) are actively
temperature-controlled between the reaction cavity (109) and the
sensor apparatus (113), and/or in that the support (113D) is
directly temperature-controlled in order to temperature-control the
capture molecules (M) and/or analytes (A) and/or amplification
products (V), and/or to reach a corresponding hybridisation
temperature (TH). 10. Method according to aspect 9, characterised
in that, after leaving the reaction cavity (109), the amplification
products (V) are brought to the hybridisation temperature (TH) in
multiple stages, and/or are actively temperature-controlled in
advance, preferably preheated, in an intermediate
temperature-control cavity (110) immediately before the sensor
apparatus (113), in particular to a temperature above the
hybridisation temperature (TH) and/or to at least 70.degree. C. or
80.degree. C. and/or at most 99.degree. C. or 95.degree. C., and/or
are subsequently temperature-controlled in or on the sensor
apparatus (113) and/or are temperature-controlled to the
corresponding hybridisation temperature (TH), in particular heated
and/or cooled. 11. Method according to aspect 9 or 10,
characterised in that different analytes (A) are amplified,
preferably in parallel and/or independently from one another and/or
in a plurality of reaction cavities (109), and/or a first group of
amplification products (V) and a second group of different
amplification products (V) are formed, preferably in parallel
and/or independently from one another and/or in different reaction
cavities (109). 12. Method according to aspect 11, characterised in
that the amplification products (V) and/or the first group and the
second group are fed to the sensor apparatus (113) in succession
and/or are bonded to the corresponding capture molecules (M) in
succession and/or are detected or determined in a single or common
detection process. 13. Method according to any of one aspects 9 to
12, characterised in that the amplification products (V) and/or the
first group and the second group are actively
temperature-controlled, preferably heated, when the fluid is
flowing through, in particular in order to denature the
amplification products (V). 14. Method according to any one of
aspects 9 to 13, characterised in that the amplification products
(V) and/or the first group and the second group are bonded to the
corresponding capture molecules (M) at different hybridisation
temperatures (TH). 15. Method according to any one of aspects 9 to
14, characterised in that the analytes (A) are amplified by means
of an amplification reaction, in particular PCR, and/or
nucleic-acid products are produced as amplification products (V)
from the analytes (A).
* * * * *