U.S. patent application number 16/842061 was filed with the patent office on 2020-07-30 for system for conducting the identification of bacteria in urine.
The applicant listed for this patent is POCARED Diagnostics LTD.. Invention is credited to William G. Atterbury, Russell H. Barnes, Douglas E. Boyd, Joseph D. Dennis, Jonathan Gurfinkel, Dave Holley, Steven E. Huckaby, Gal Ingber, Thomas A. Klausing, John S. Laudo, Kevin Sadeski, Jason A. Schaefer, K. Bryan Scott, Carol Stillman, Sherwood Talbert, John Tallarico.
Application Number | 20200240977 16/842061 |
Document ID | 20200240977 / US20200240977 |
Family ID | 1000004750168 |
Filed Date | 2020-07-30 |
Patent Application | download [pdf] |
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United States Patent
Application |
20200240977 |
Kind Code |
A1 |
Ingber; Gal ; et
al. |
July 30, 2020 |
System for Conducting the Identification of Bacteria in Urine
Abstract
A system for conducting the identification and quantification of
micro-organisms, e.g., bacteria in urine samples which includes: 1)
several disposable cartridges for holding four disposable
components including a centrifuge tube, a pipette tip having a 1 ml
volume, a second pipette tip having a 0.5 ml volume, and an optical
cup or cuvette; 2) a sample processor for receiving the disposable
cartridges and processing the urine samples including transferring
the processed urine sample to the optical cups; and 3) an optical
analyzer for receiving the disposable cartridges and configured to
analyze the type and quantity of micro-organisms in the urine
sample. The disposable cartridges with their components including
the optical cups or cuvettes are used in the sample processor, and
the optical cups or cuvettes containing the processed urine samples
are used in the optical analyzer for identifying and quantifying
the type of micro-organism existing in the processed urine
samples.
Inventors: |
Ingber; Gal; (Oranit,
IL) ; Atterbury; William G.; (Columbus, OH) ;
Barnes; Russell H.; (Columbus, OH) ; Boyd; Douglas
E.; (Columbus, OH) ; Dennis; Joseph D.;
(Granview Heights, OH) ; Gurfinkel; Jonathan;
(Omer, IL) ; Holley; Dave; (Lancaster, OH)
; Huckaby; Steven E.; (Blacklick, OH) ; Klausing;
Thomas A.; (Powell, OH) ; Laudo; John S.;
(Columbus, OH) ; Sadeski; Kevin; (Canal
Winchester, OH) ; Schaefer; Jason A.; (Coral Springs,
FL) ; Scott; K. Bryan; (Westerville, OH) ;
Stillman; Carol; (Powell, OH) ; Talbert;
Sherwood; (Columbus, OH) ; Tallarico; John;
(Powell, OH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
POCARED Diagnostics LTD. |
Rehovot |
|
IL |
|
|
Family ID: |
1000004750168 |
Appl. No.: |
16/842061 |
Filed: |
April 7, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15425253 |
Feb 6, 2017 |
10656140 |
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16842061 |
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14330450 |
Jul 14, 2014 |
9606105 |
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15425253 |
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12682402 |
Sep 15, 2010 |
8808649 |
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PCT/US2008/079533 |
Oct 10, 2008 |
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14330450 |
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60978810 |
Oct 10, 2007 |
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60978820 |
Oct 10, 2007 |
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60978856 |
Oct 10, 2007 |
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60978862 |
Oct 10, 2007 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C12Q 1/04 20130101; G01N
2035/0458 20130101; G01N 35/025 20130101; G01N 2021/6482 20130101;
G01N 2201/0415 20130101; G01N 35/04 20130101; G01N 2035/0448
20130101; G01N 21/0303 20130101; G01N 2035/0444 20130101; G01N
33/493 20130101; G01N 2035/0436 20130101 |
International
Class: |
G01N 33/493 20060101
G01N033/493; C12Q 1/04 20060101 C12Q001/04; G01N 21/03 20060101
G01N021/03; G01N 35/02 20060101 G01N035/02; G01N 35/04 20060101
G01N035/04 |
Claims
1-16. (canceled)
17. A method for processing a fluid sample in preparation for the
identification and quantification of micro-organisms in the fluid
sample, the steps comprising: a) obtaining a fluid sample; b)
passing the fluid sample through a filter; c) obtaining a sample of
the filtered fluid sample and placing it into a centrifuge tube; d)
obtaining a dilution of the dissolved material in the fluid sample
while retaining bacteria in the fluid sample by performing the
following steps: 1) centrifuging the sample of step c); 2)
decanting a portion of the fluid in the centrifuge tube of step 1);
3) replacing the decanted solution of step 2) with a saline
solution; and e) transferring the final solution of step 4) into an
optical cup or cuvette for analysis.
18. The method of claim 17, wherein the fluid sample is urine.
19. The method of claim 17, wherein step d) results in about a
1,000,000:1 dilution.
20. The method of claim 17, wherein the sample of step (d)(1) is
centrifuged at about 12,000 g-forces.
21. The method of claim 17, wherein about 95% of the fluid from
step (d)(2) is decanted.
22. The method of claim 17, wherein after step d(3), further
comprising a step of repeating steps (1), (2) and (3) and then
transferring the final solution into an optical cup or cuvette for
analysis.
23. The method of claim 17, wherein after step (d)(3), further
comprising a step of repeating steps (1), (2) and (3) for a
plurality of times and then transferring the final solution into an
optical cup or cuvette for analysis.
24. The method of claim 23, wherein the plurality of times is at
least five times.
25-37. (canceled)
38. A method for identifying the type and quantity of
micro-organisms in a fluid sample, the steps comprising: a)
obtaining a fluid sample; b) passing the fluid sample through a
filter; c) obtaining a sample of the filtered fluid and placing it
into a centrifuge tube; d) obtaining a dilution of the dissolved
material in the urine sample while retaining bacteria in the urine
sample by performing the following steps: 1) centrifuging the
sample of step c); 2) decanting a portion of the fluid in the
centrifuge tube of step 1); 3) replacing the decanted solution of
step 2) with a saline solution; and e) transferring the final
solution of step 4) into an optical cups or cuvette; and f)
subjecting the optical cup or cuvette to an optical analysis having
optics which include exciting the fluid sample with a plurality of
different wavelengths, collecting and detecting the fluorescent
emissions; and directing the fluorescent emissions into a
spectrometer.
39. The method of claim 38, wherein step d) results in about a
1,000,000:1 dilution.
40. The method of claim 38, wherein the sample of step (d)(1) is
centrifuged at about 12,000 g-forces.
41. The method of claim 38, wherein about 95% of the fluid from
step (d)(2) is decanted.
42. The method of claim 38, wherein after step d(3), further
comprising a step of repeating steps (1), (2) and (3) and then
transferring the final solution into an optical cup or cuvette for
analysis.
43. The method of claim 38 wherein after step (d)(3), further
comprising a step of repeating steps (1), (2) and (3) for a
plurality of times and then transferring the final solution into an
optical cup or cuvette for analysis.
44. The method of claim 43 wherein the plurality of times is at
least five times.
45. A method for identifying the type and quantity of
micro-organisms in a fluid sample, the steps comprising: a)
obtaining a fluid sample; b) passing the fluid sample through a
filter; c) obtaining a sample of the filtered fluid and placing it
into a removable tube; and d) transferring the sample of the
filtered fluid into an optical cup or cuvette, said optical cup or
cuvette comprising a container having an open portion and a
sidewall that is continuously tapered along their length to a
truncated conical end and a reflective surface; and e) subjecting
the optical cup or cuvette to an optical analysis having optics
which include exciting the fluid sample with a plurality of
different wavelengths, collecting and detecting the fluorescent
emissions, and directing the fluorescent emissions into a
spectrometer, wherein during the optical analysis, a light is
directed down a middle portion of the container through the open
portion of the container and into the fluid sample, wherein the
sidewall is continuously tapered along its length to a truncated
conical end configured to assist with the optical analysis and the
reflective surface of the container reflects fluorescent emissions
back in an upwardly direction out of the fluid sample and through
the open portion of the container to an optics system for the
optical analysis.
46. The method of claim 45, wherein the container includes a lip
for suspending the container from a top surface of a cartridge.
47. The method of claim 46, wherein the cartridge includes a body
and the container is located within an opening in the cartridge
body.
48. The method of 45, wherein the container has a flat bottom
surface.
49. The method of claim 45, wherein the reflective surface is a
coated onto the container or is a separately formed layer of
material.
50. The optical cup or cuvette of claim 45, wherein the reflective
surface is an inner reflective surface of the container or is
incorporated onto the container material.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims priority as a divisional
application of U.S. patent application Ser. No. 14/330,450 entitled
"System for Conducting the Identification of Bacteria in Urine"
filed Jul. 14, 2014, which claims priority to U.S. patent
application Ser. No. 12/682,402 entitled "System For Conducting The
Identification Of Bacteria In Urine" filed Apr. 9, 2010, which is
the United States national phase of International Patent
Application No. PCT/US2008/079533 entitled "System For Conducting
The Identification Of Bacteria In Urine" filed Oct. 10, 2008, which
claims priority to U.S. Provisional Application Nos. 60/978,810,
60/978,820, 60/978,856 and 60/978,862, all filed on Oct. 10, 2007,
which are herein incorporated by reference in their entirety.
BACKGROUND OF THE INVENTION
Field of the Invention
[0002] The present invention relates to a system for conducting the
identification and quantification of micro-organisms, e.g.,
bacteria in urine samples. More particularly, the invention relates
to a system for identifying bacteria in urine samples which
includes: 1) a disposable cartridge or holder for holding
disposable components including a centrifuge tube, two pipette tips
with a different volume capacity, and an optical cup or cuvette; 2)
a sample processor for processing or preparing the urine samples;
and 3) an optical analyzer for analyzing the processed urine
samples. The disposable cartridge with its four components is used
in the sample processor and the optical cup or cuvette in
particular is used in the optical analyzer.
Description of Related Art
[0003] In general, current-day practice for identifying
micro-organisms, e.g., bacteria in urine samples involves a
complex, lengthy and expensive process for identifying and
specifying micro-organisms in microbiology labs. In the current
process, the samples are accepted into the lab. These specimens are
then sorted and labeled and then they are inoculated onto blood
agar medium using sterilized loop. The specimens are then inserted
into a dedicated incubator for a 24-hour period. A day later, the
lab technicians screen the specimens for positive and negative
cultures. In general, most of the cultures are negative and they
are manually reported. The organisms for the positive cultures are
isolated and suspended in a biochemical fluid. This involves
suspension, dilution, vortexing and turbidity measurements
resulting in biochemical waste products. The cultures are then
subjected to a species identification and antibiotics
susceptibility testing exposing the suspensions to multiple
reagents. After another 6 to 24-hour incubation period, the
findings are interpreted and reported by lab technicians. This
entire process generally takes 11 steps and 50 hours to obtain
specimen results and the process is labor intensive.
[0004] Commonly owned U.S. Publication No. US 2007/0037135 A1, the
contents of which are herein incorporated by reference, discloses a
system for identifying and quantification of a biological sample
suspended in a liquid. Paragraph [0038] of this publication states
that sample cuvettes are used. These cuvettes are said to be well
known in the art and are typically square or rectangular in shape
(having a well area to contain the sample) and are made of a
transparent material such as glass or a polymeric material.
[0005] There is a need, therefore, particularly for species
identification of the above lab procedure to provide a more
efficient, and less time consuming process which requires less
labor.
SUMMARY OF THE INVENTION
[0006] The system of the invention streamlines this current system
for obtaining specimen results. The system is environmentally
friendly, enables a rapid diagnosis, results are consistent, no
reagents are needed and there is a multifunctional diagnosis. Urine
samples are contained within disposable cartridges which hold four
disposable components, i.e., a centrifuge, two pipette tips with a
different volume and an optical cuvette. The cartridges are bar
coded and tied in with the patient's ID. The cartridges are
inserted in a magazine which is then inserted into a sample
processor which processes the specimens. The prepared specimens are
transferred into the optical cuvettes and then the magazine is
inserted into an optical analyzer which analyses the specimens. The
optical analyzer analyses and generates the complete results
enabling ultimate treatment of the bacteria. The system does not
require a sophisticated operator and gives rapid results. The
system increases efficiency, improves workload, saves time and
money and is easy to operate. The sample preparation can be
performed in parallel with the specimen analysis process and from 1
to 50 specimens can be analyzed simultaneously.
[0007] The system includes a plurality of disposable cartridges for
holding a plurality of disposable components including a centrifuge
tube, a first pipette tip with a 1 ml volume; an optical urine
sample cuvette, and a second pipette tip with a 0.5 ml volume; a
sample processor for receiving the plurality of disposable
cartridges and configured to process and prepare the urine sample
of each disposable cartridge and to transfer the urine samples into
the respective optical cuvette of each of the disposable
cartridges; and an optical analyzer for receiving the cartridge
with the optical cuvettes containing the processed urine samples
and analyzing and generating the specimen results. The entire
process of processing the urine specimens in the sample processor
and analyzing them in the optical analyzer takes about 20 minutes
for a single specimen and up to 2 hours for 50 specimens.
[0008] The disposable cartridge and the disposable components
increase efficiency, improve workload and save time and money since
the components necessary for the preparation or processing of the
urine samples are conveniently located in one place, i.e., in a
cartridge. Additionally, less manpower or manual handling of the
components is required for the processing/analyzing of the urine
samples. There is also the added convenience in that the cartridge
and its components are disposable. That is, these items do not need
to be sterilized for the next urine specimen identification process
and contamination of the work area and/or surrounding environment
is minimized.
[0009] The sample processor of the invention streamlines the
current practice for processing urine samples for analysis. The
sample processor of the invention is automated, fully compact,
self-contained and does not require any reagents. Urine samples are
injected into centrifuge tubes which are carried in removable
cartridges. The specimens are bar coded and tied with the
cartridges which are also bar coded. The cartridges are mounted in
a magazine which is then inserted into the sample processor which
processes the specimens in preparation for analysis. The sample
processor does not require a sophisticated operator and rapidly
processes the samples or specimens. The sample processor increases
efficiency, improves workload, saves time and money and is easy to
operate. The processing of several samples can be performed in
about 20 minutes for a single specimen and up to 1 hour for about
50 specimens.
[0010] The optical analyzer contains an optics system, a thermal
control and a drawer which has a rotatable table for receiving,
supporting and rotating a magazine containing a plurality of
disposable cartridges with optical cups or cuvettes which contain
the urine samples to be analyzed. The optical analyzer also
contains a bar code reader for inventorying the urine samples and a
level sensor that verifies that each optical cup or cuvette
contains the correct volume of processed urine sample. When the
drawer with the magazine is inserted into the optical analyzer, the
drive mechanism for the rotatable table supporting the magazine
rotates and registers the magazine relative to the bar code reader
and then rotates and registers the magazine relative to the optics
system. The optics system includes an excitation module unit, an
optical collection unit and a spectrometer. The temperature of each
cup or cuvette is decreased to a temperature which will slow the
metabolism of the bacteria in the urine samples while increasing
the fluorescence signal. A thermal control cools a large thermal
mass which is located on the rotatable table underneath the
magazine containing the disposable cartridges with urine sample
cups or cuvettes. An infrared temperature sensor detects and
monitors the temperature of each urine sample.
[0011] A related method for identifying the type of micro-organism
and quantifying it in a urine sample includes the steps of
obtaining a urine sample; passing the urine sample through a ten
micron filter; obtaining a 2 ml sample of the filtered urine and
placing it into a centrifuge tube; obtaining a 1,000,000:1 dilution
of the dissolved materials in the urine retaining bacteria in the
urine sample by centrifuging the 2 ml sample at about a 12,000
g-force, decanting about 95% of the fluid in the centrifuge tube,
replacing the decanted solution with a saline solution, and
repeating these steps about five times; transferring the final
solution into an optical cup or cuvette; and subjecting the optical
cup or cuvette to an optical analysis having optics which include
exciting the urine sample with at least five different wavelengths,
collecting and detecting the fluorescent emissions; and directing
the fluorescent emissions into a spectrometer.
[0012] These and other objects and advantages of the invention will
be made apparent from the following description taken together with
the drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0013] FIG. 1A is a top perspective view of a magazine having a
plurality of disposable cartridges.
[0014] FIG. 1B is a top perspective view of a disposable cartridge
used in the magazine shown in FIG. 1A.
[0015] FIG. 2 is a front sectional view illustrating the components
of the disposable cartridge of FIG. 1 in phantom.
[0016] FIG. 3A is a perspective view of a sample processor
illustrating in phantom the several components of the sample
processor of the system of the invention.
[0017] FIG. 3B is an additional perspective view of a sample
processor illustrating in phantom the several components of the
sample processor of the system of the invention.
[0018] FIG. 4A is a perspective view of an optical analyzer
illustrating in phantom the several components of the optical
analyzer of the system of the invention.
[0019] FIG. 4B is a perspective view of an optics system
illustrating in phantom the several components of the optics of the
system of the invention.
[0020] FIG. 4C is an additional perspective view of an optical
analyzer illustrating in phantom the several components of the
optical analyzer of the system of the invention.
[0021] FIG. 5 is a schematic illustrating mirrored convex "horn"
that may be provided at the entrance of a slit of a
spectrometer.
[0022] FIG. 6 is a perspective view of a centrifuge illustrating in
phantom the several components of the centrifuge of the system of
the invention.
[0023] FIG. 7 is an additional perspective view of a sample
processor illustrating in phantom the several components of the
sample processor of the system of the invention.
DETAILED DESCRIPTION OF THE INVENTION
[0024] The present invention will be described with reference to
the accompanying drawings where like reference numbers correspond
to like elements.
[0025] Referring to FIGS. 1-4, the system for conducting the
identification of bacteria in urine samples includes a disposable
cartridge 12 (FIGS. 1 and 2); a sample processor 14 (FIGS. 3A, 3B,
6 and 7); and an optical analyzer 16 (FIGS. 4A, 4B, and 4C). As
shown in FIGS. 1 and 2, cartridge 12 contains four disposable
components, which are a centrifuge tube 18, a first pipette tip 20
having a 1 ml volume, an optical cup or cuvette 22, and a second
pipette tip 24 having a 0.5 ml volume. It is to be understood that
the presently described inventive system is appropriate for the
identification of bacteria in any fluid and is not limited to
bacteria samples contained in urine.
[0026] The centrifuge tube 18 is a container that has an elongated
body 18b with a tapered end indicated at 18a. In general, the
centrifuge tube 18 initially contains the urine sample and the
first pipette tip 20 may be used to dilute the urine-dissolved
constitutes, and the second pipette tip 24 may be used to transfer
the diluted urine sample into the optical cup or cuvette 22 for
optical analysis. The disposable cartridge 12 and its disposable
components 18, 20, 22, and 24 may be made of a plastic material
which is easily molded and inexpensive to manufacture.
[0027] Still referring to FIG. 2, the disposable components 18, 20,
22, and 24 are each contained within separate locations 30, 32, 34,
and 36, respectively of the disposable cartridge 12. As is shown,
the bottom of compartment 32 which receives and carries the first
pipette tip 20 is closed so that any drip from the first pipette
tip 20 will not contaminate the surface below the disposable
cartridge 12. Each component 18, 20, 22, and 24 is suspended within
its respective location 30, 32, 34, and 36 via a lip 40, 42, 46,
and 48, respectively attached to each component 18, 20, 22, and 24
which is supported by the top surface 50 of disposable cartridge
12.
[0028] Referring to FIGS. 2 and 4A, an optical cup or cuvette 22
may be used in the optical analyzer 16 of FIG. 4A. Preferably, the
urine samples are prepared with a saline solution since saline
solutions minimize background fluorescence while maintaining the
integrity of the bacteria which particularly is important when
using optics in the urine analysis process. The optical cup or
cuvette 22 will include a reflective coating to assist in the
optical analysis. The optical cup or cuvette 22 may be made of an
ABS plastic material, glass or a metallic material, e.g., aluminum,
and then coated with or layered with the reflective material.
Alternatively, in the manufacturing of the optical cup or cuvette
22, the layer of reflective material may be incorporated onto the
plastic, glass or metallic material. As best shown in FIG. 2, the
optical cup or cuvette 22 includes a tapered end indicated at 22a
in order to assist with the optical analysis. It is anticipated
that the UV light source in the optical analyzer 16 (FIGS. 4A, 4B
and 4C) be directed down the middle of the cup or cuvette 22 for
the optical analysis of the urine specimen in the cup or cuvette
22.
[0029] Several disposable cartridges 12 each containing the four
disposable components 18, 20, 22, and 24 are then inserted into a
magazine 26 shown at the top of FIG. 1, which is then loaded into
the sample processor 14 as shown in FIG. 3. Magazine 26 contains
several disposable cartridges 12 some of which are numbered, each
cartridge 12 has a unique bar code as indicated at 28 in FIG. 1
that is paired with the specimen of a patient. Alternatively, the
magazine 26 can then be inserted into a device for the optical
analysis of the urine samples. Preferably, the same magazine 26
used in obtaining processed urine samples in a sample processor is
used in the device for the optical analysis of the processed urine
samples.
[0030] The sample processor 14 of FIGS. 3A and 3B contains a
centrifuge 30, a rotatable gripper mechanism 32 which picks up the
centrifuge tube 18 (FIG. 1) of each disposable cartridge 12 and
inserts the centrifuge tube 18 into the centrifuge 30; two movable
fluid transfer arms 34, 34a which are used to dilute the dissolved
material in the urine samples via the pipette tip 20 (FIGS. 1 and
2) and to transfer the diluted sample to the optical cup or cuvette
22 (FIG. 2) via the pipette tip 24; and a syringe pump dispenser
fluid system 36 for delivering water to the samples for dilution
purposes. The sample processor 14 also includes a drawer 38 which
has a rotatable table 40 which receives, supports, and rotates the
magazine 26 when the drawer 38 is inserted into the sample
processor 14. The drawer 38 contains a magazine drive mechanism
(not shown) which rotates the magazine 26.
[0031] In general, centrifuge tube 18 contains about 2 ml sample of
filtered urine which is placed into the centrifuge tube by the
user. This sample may then be sufficiently diluted with a saline
solution or water by centrifuging the sample followed by using the
first pipette tip 20 with the 1.0 ml volume to decant the
supernates in two decant cycles followed by refilling of the
centrifuge tube 18 with a saline or water. The second pipette tip
24 having the 0.5 ml volume may then be used to draw out about 500
.mu.l of fluid from centrifuge tube 18 and then to dispense this
500 .mu.l of fluid into the respective optical cup or cuvette 22 of
the designated patient. This second pipette tip 24 can then be
inserted into the first pipette tip 20 and both pipette tips 20, 24
can be disposed of properly. It is believed that one pipette tip
may be used to dilute and draw out instead of two pipette tips.
This process may be done manually or may be done automatically.
[0032] The loading and unloading of the magazine 26 is accomplished
with the several disposable cartridges 12 mounted on the rotatable
table 40 (FIG. 1 and FIG. 2). The manual drawer contains a magazine
drive mechanism (not shown). Once the magazine 26 is inserted into
the sample processor 14, the drive mechanism (not shown) for
rotatable table 40 rotates the magazine 26; the bar code reader
(element 58 in FIG. 4A) inventories the samples, a level sensor
(not shown) verifies that samples were dosed properly; and a second
sensor (not shown) verifies that all of the necessary disposable
components 18, 20, 22, and 24 (FIG. 2) are contained in each
disposable cartridge 12.
[0033] The transfer of the centrifuge tube 18 (FIG. 2) into the
centrifuge 30 (FIGS. 3A and 3B) will now be described. A centrifuge
lid 30a of the centrifuge 30 is oriented to allow the rotatable
gripper mechanism unit 32 to access and load the centrifuge 30. The
drive mechanism of the rotatable table 40 is configured to align
the centrifuge tube 18 of each disposable cartridge 12 into
position relative to the rotatable unit 32. The gripper 32a of
rotatable gripper mechanism 32 selects the centrifuge tube 18 for
transfer from the magazine 26 and into the centrifuge 30. The
centrifuge rotor (not shown) is configured to align a vacant
centrifuge holder of centrifuge 30 in the load position. The
gripper 32a referred to as a "Theta Z gripper" is a radial member
that rotates and has a downward and upward movement for picking up
and setting a centrifuge tube 18 into a vacant centrifuge holder of
centrifuge 30. The lid of centrifuge 30 is closed after all of the
centrifuge tubes 18 are placed into the centrifuge 30.
[0034] Centrifuge 30 (FIG. 6) is automatically operated to spin the
centrifuge tubes 18 at about a 12,000 g-force for about 2 minutes.
The centrifuge 30 includes tube holders that are configured to
rotate each of the centrifuge tubes 18 about ninety degrees upon
rotation of the centrifuge 30. The centrifuge allows for precise
positioning and position tracking so that correct tubes are
returned to cartridges in the magazine after centrifugation. This
action results in the solid formation of the bacteria present in
the urine sample at the bottom of the centrifuge tube 18.
[0035] There are two fluid transfer arms 34, 34a (FIGS. 3A and 3B)
for removing the supernates from two samples of two disposable
cartridges 12 at a time. After the two fluid transfer arms 34, 34a
(FIGS. 3A and 3B) obtain the pipette tip 20 (FIG. 2) with a 1 ml
volume, each of the fluid transfer arms 34 and 34a (FIGS. 3A and
3B) makes two consecutive trips to the centrifuge tube 18, each
time drawing fluid from the tube 18 and dispensing this fluid into
a waste port (not shown) of sample processor 14 before returning
the pipette tip 20 to its location on the disposable cartridge that
is being sampled and before continuing with the next sample in the
disposable cartridge 12 that is rotated to be registered in the
sampling location of sample processor 14.
[0036] The syringe pump dispenser fluid system 36, is illustrated
in FIG. 7, for delivering water or saline to the samples for
dilution purposes. The waste fluid which had been decanted from a
centrifuge tube 18 as described in the preceding paragraph is
replaced with clean process fluid via system 36. Two syringe pumps
dispense this clean process fluid into the centrifuge tube 18 from
which the waste fluid had been removed in the previous step. During
the final refill step, a smaller amount of clean fluid is used in
order to get the bacteria level in the cartridge tube 18 to the
required concentration.
[0037] After the sample in centrifuge tube 18 has been sufficiently
diluted with the clean fluid, one of the two fluid transfer arms
34, 34a (FIGS. 3A and 3B) transfers the processed sample in
centrifuge tube 18 to the optical cup or cuvette 22 of its
respective disposable cartridge 12. One of the fluid transfer arms
34, 34a grasps the pipette tip 24 having the 0.5 ml volume, which
until now has not been used in this process. This pipette tip 24
with the smaller volume is used to draw out about 500 .mu.l of
fluid from centrifuge tube 18 and is used to dispense this fluid
into the respective optical cup or cuvette 22 of the designated
patient. This pipette tip 24 with the smaller volume is then
inserted into the pipette tip 20 with the larger volume via the
fluid transfer arm 34 or 34a for disposal of both pipette tips 20,
24.
[0038] The metering/decanting, metering/refilling, and
metering/fluid transferring process described herein above is to
obtain preferably, approximately a 1,000,000:1 dilution of the
dissolved materials retaining bacteria in the urine sample in
centrifuge tube 18. This can be achieved by 1) centrifuging,
through means known to those skilled in the art, the urine sample
at a 12,000 g-force; 2) decanting about 95% of the fluid by using
the first pipette tip 20; 3) replacing the decanted solution of 2)
with a saline solution; and 4) repeating steps 1), 2), and 3) at
least five times by using the first pipette 20. The final processed
urine sample in centrifuge tube 18 can then be decanted via the
second pipette 24 into the optical cup or cuvette 22.
[0039] The final processed urine sample in optical cup or cuvette
22 can then be used in an optical analysis for determining the
micro-organism's identity and/or quantity in the urine sample in
optical cup or cuvette 22. This information can be obtained by
using the system as disclosed in the aforesaid U.S. Publication No.
2007/0037135 A1.
[0040] Each of the steps described above for one centrifuge tube 18
is done in the sample processor 14 for each of the disposable
cartridges 12 in magazine 26. It is to be appreciated that the
waste fluid of each disposable cartridge 12 is disposed into a
receptacle (not shown) in sample processor 14 or is plumbed
directly into a drain. The waste disposables, i.e., the disposable
cartridge 12 and disposable components 18, 20, 22, and 24 remain on
the magazine 26 for manual removal when the magazine 26 is unloaded
in preparation for the next operation of the sample processor 14
for processing the next batch of urine samples.
[0041] The following steps are involved in processing the urine
samples in preparation for analysis via the optical analyzer 16 of
FIGS. 4A, 4B, and 4C. In general, a sample of urine is obtained in
a test tube. This sample is passed through a ten micron filter from
which a 2 ml sample is obtained and placed into the centrifuge tube
18. The desired diluted sample, i.e., 1,000,000:1 dilution of
dissolved materials while retaining bacteria in the urine sample is
obtained by centrifuging this 2 ml sample at about a 12,000
g-force; and decanting 95% of the fluid. This latter step is
repeated five times wherein the decanted solution is replaced each
time with a saline solution. A saline solution is selected for this
process in that it minimizes background fluorescence which comes
into play when the processed urine sample is inserted into the
optical analyzer 16 while maintaining the bacteria integrity.
[0042] The optical analyzer 16 of FIGS. 4A, 4B, and 4C will now be
described. With reference to FIG. 4A, the optical analyzer 16
includes an optics system 44 (shown in greater detail in FIGS. 4B
and 4C), a thermal control unit (not shown), a drawer 50 which has
a rotatable table 52 which receives, supports, and rotates a
magazine 54 containing a plurality of holders 56 for receiving the
disposable cartridges 12 in which cups or cuvettes 22 contain the
processed urine samples which are to be analyzed, and a bar code
reader 58 (FIG. 4A).
[0043] As can be appreciated, a cartridge 12 that has the cups or
cuvettes 22 containing the processed urine sample for optical
analysis are placed into the holders 56 of the magazine 54. FIG. 4A
illustrates the magazine 54 mounted on the rotatable table 52 being
loaded into the optical analyzer 16. Drawer 50 is pulled out
manually for the loading and unloading of magazine 54. Drawer 50
contains the thermal control unit (not shown) and a drive mechanism
(not shown). Alignment features on the magazine 54 and drawer 50
allow the operator to orient the magazine 54 properly on the drive
mechanism and the thermal control unit when the magazine 54 is
loaded onto the rotatable table 52. Once the drawer 50 and magazine
54 are manually inserted into the optical analyzer 16, the drive
mechanism rotates the magazine 54 at which time a bar code reader
station 58 (FIG. 4A) inventories the samples. A level sensor (not
shown) verifies that each optical cup or cuvette 22 contains the
correct sample volume. An operator can access the optical analyzer
16 when a user interface indicates that all the samples in the cups
or cuvettes 22 have been analyzed and drawer 50 is prevented from
being opened when any of the components of optical analyzer 16 are
moving or when the UV light sources of the optics system 44 are
on.
[0044] FIG. 4A illustrates the magazine 54 on rotatable table 52
while being positioned within optical analyzer 16. The optical
analyzer 16 further includes a mechanical locking system (not
shown) which positions the drawer 50 accurately with respect to the
optics system 44. The drive mechanism is configured to
automatically rotate the magazine 54 to position each cartridge 12
into the bar code reader station 58 and into precise alignment with
the optics system 44. A second mechanical locking system (not
shown) is used to secure each cup or cuvette 22 in its proper
positioning relative to the optics system 44 for optical
analysis.
[0045] FIG. 4A illustrates the thermal control for the optical cups
or cuvettes 22. Preferably, the temperature of each cup or cuvette
22 is decreased to a temperature which will slow the metabolism of
the bacteria while increasing the fluorescence signal. The thermal
control unit 46 which is a thermal electric cooler (TEC) cools a
large thermal mass 60 which is located on the rotatable table 52
underneath the magazine 54. The thermal mass 60 (FIG. 4A) is in
direct contact with the optical cups or cuvettes 22, and an
infrared temperature sensor (not shown) detects and monitors the
temperature of each sample prior to the sample being rotated and
positioned within the optics system 44.
[0046] The optics system 44 of the optical analyzer 16 will now be
described. The optics system is shown in greater detail in FIG. 4B.
The optics system 44 contains three separate units, that is, an
excitation unit 44(a), an optical collection unit 44(b) and a
spectrometer. Excitation will be provided by a ultra-violet (UV)
light source, which preferably will be LED (light emitting diode).
A series of five LED modules provide an excitation unit 44(a) will
sequentially provide excitation signals to each sample cup or
cuvette 22 at five different excitation wavelengths which will be
applied to each sample cup or cuvette 22 in the same order. The
excitation time will be approximately 14 seconds per wavelength.
The excitation emissions are directed via lenses and filters 44(d)
to be directed an upper surface of the sample in the cuvette 22 as
shown in FIG. 2. In order to narrow or control the shape of each
excitation wavelength, narrow bandwidth filters will be used. These
filters will direct in a downwardly direction the excitation
wavelengths E to the sample cups or cuvettes 22 and the fluorescent
emissions F will be reflected back in an upwardly direction to the
optical collection unit from the same position of the cassette. The
fluorescent emissions can be separated and directed via a filter
arrangement. FIG. 4C illustrates the positioning of the optics
system 44. As described previously, mechanical locking features
position the drive mechanism such that the sample cup or cuvette 22
is aligned precisely. This precise alignment allows for the
reflection of the fluorescent emission to the optics system 44
allowing for measurement of fluorescence. Optical elements (not
shown) are utilized to gather and direct the fluorescent emissions
into the spectrophotometer for measurement.
[0047] In addition, the optical collection unit includes optical
elements to gather and direct the fluorescent emissions of the
samples in the cups or cuvettes 22 into the spectrometer.
[0048] The optics system 44 may include a Czerny-Turner
spectrometer with a CCD (charged couple device) Photon Detector,
whereby fluorescent photons are reflected by several mirrors before
contacting the CCD device. The emitted fluorescence will be
monitored on the CCD device by integrating for a period of time. It
is also envisioned that the Czerny-Turner spectrometer be modified
with additional cylindrical lenses adjacent the entrance slit and
the CCD device in order to improve photon usage efficiency.
Additionally, as schematically illustrated in FIG. 5, mirrored
convex "horn" H may be provided at the entrance of the slit S of
the spectrometer SM to direct additional photons through the slit
S.
[0049] Referring to FIG. 4A, the optics system 44 will include a
light-tight enclosure or housing 64 in order to minimize light
entering the optics system 44, and the camera of the CCD device
will include a thermal electric cooler (TEC) (not shown) for
transferring heat from the camera chip to the enclosure or housing
64 of the optics system 44.
[0050] Both the sample processor 14 and the optical analyzer 16
will have a HEPA air-filtering system for ventilation purposes in
filtering the air entering and exiting the sample processor 14 and
the optical analyzer 16. The sample processor 14 and the optical
analyzer 16 will also have a slight positive pressure during
operation. However, this pressure will decrease to atmospheric
pressure when the user is allowed to open the door. Power to both
the sample processor 14 and the optical analyzer 16 will be
supplied from a separate rack mounted power supply module.
[0051] It is further envisioned that the LED intensity will be
monitored to correlate the emitted fluorescence with the intensity
of the excitation fluorescence. In particular, the information
obtained by the optical analyzer 16 may be used to generate graphs
similar to FIGS. 5 through 9 of U.S. Publication No. 2007/0037135
A1, described in greater detail below. The graphs represent for the
concentration of the bacteria in the sample cups or cuvettes 22,
the fluorescence intensity, the emission wavelengths and the
excitation wavelengths.
[0052] As discussed hereinabove, the optical analyzer 16 provides
results that are then used to identify the type of bacteria in the
urine samples. This can be done by coupling the optical analyzer 16
to a computer module (not shown) and feeding in the acquired
information of the optical analyzer 16, such as the fluorescence
emission, into the computer module. The computer module may perform
multivariate analysis on the fluorescence excitation-emission
matrices of the urine samples to identify and quantify the urine
samples in a manner similar to that disclosed in the above U.S.
Publication No. US 2007/0037135 A1. Here, the system includes a
fluorescence excitation module which includes an excitation light
source, a sample interface module for positioning the sample to
receive the light source, a fluorescence emission module and a
detection device. The computer module described above is coupled to
the fluorescence module. The multivariate analysis may comprise
extended partial least squared analysis for identification and
quantification of the urine samples.
[0053] It is still further envisioned that a "homogenitor tube"
will be used to mix the different LED packages output into a
uniform UV light source. A typical "homogenitor tube" for use in
the invention will be similar to that known to those skilled in the
art.
[0054] The present invention has been described with reference to
the preferred embodiments. Obvious modifications and alterations
will occur to others upon reading and understanding the preceding
detailed description. It is intended that the invention be
construed as including all such modifications and alterations.
* * * * *