U.S. patent application number 10/826994 was filed with the patent office on 2008-08-28 for blood coagulation test cartridge, system, and method.
Invention is credited to Daniel S. Cheek, Cynthia T. Clague, Douglas D. Nippoldt.
Application Number | 20080206880 10/826994 |
Document ID | / |
Family ID | 34965011 |
Filed Date | 2008-08-28 |
United States Patent
Application |
20080206880 |
Kind Code |
A9 |
Clague; Cynthia T. ; et
al. |
August 28, 2008 |
BLOOD COAGULATION TEST CARTRIDGE, SYSTEM, AND METHOD
Abstract
A system and method for determining a coagulation time, e.g.,
TT, PT, aPTT, and ACT, of a test sample deposited in a test
cartridge is disclosed. A cartridge housing having upper and lower
major sides and a minor sidewall encloses a test chamber having a
test chamber pivot element and is provided with a cartridge port
for introducing a test sample into the test chamber,. Ferromagnetic
agitator vane leaflets extend from an agitator pivot element
supported by the test chamber pivot element intermediate the upper
and lower major sides for rotational motion. The agitator vane
leaflets can be swept, in response to an external magnetic field,
through the test sample in the absence of coagulation. A timer is
started when the agitator movement is commenced whereupon the
agitator moves freely. Resistance to agitator movement due to
coagulation is detected, and the coagulation time is measured.
Inventors: |
Clague; Cynthia T.;
(Minnetonka, MN) ; Cheek; Daniel S.; (Plymouth,
MN) ; Nippoldt; Douglas D.; (Oakdale, MN) |
Correspondence
Address: |
MEDTRONIC, INC.
710 MEDTRONIC PARKWAY NE
MINNEAPOLIS
MN
55432-9924
US
|
Prior
Publication: |
|
Document Identifier |
Publication Date |
|
US 20050233460 A1 |
October 20, 2005 |
|
|
Family ID: |
34965011 |
Appl. No.: |
10/826994 |
Filed: |
April 19, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60548438 |
Feb 27, 2004 |
|
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Current U.S.
Class: |
436/69 ; 422/430;
422/68.1 |
Current CPC
Class: |
G01N 15/042 20130101;
Y10T 436/111666 20150115; G01N 11/162 20130101; G01N 11/14
20130101; G01N 33/4905 20130101; G01N 2015/0092 20130101; G01N
15/05 20130101 |
Class at
Publication: |
436/069 ;
422/058; 422/068.1 |
International
Class: |
G01N 33/86 20060101
G01N033/86 |
Claims
1. A system that performs a test on a test sample determining a
coagulation test time comprising: a test cartridge comprising: a
cartridge housing having at least one cylindrical test chamber
therein into which a test sample is deposited to be tested, the
test chamber having a test chamber axis, a test chamber diameter
and a test chamber height defining a test chamber volume; and an
agitator mounted at a pivot point at the test chamber axis, the
agitator having an agitator vane adapted to be swept about the
pivot point and through the test sample in the test chamber, the
agitator vane having an agitator vane height extending
substantially through the test chamber height and an agitator vane
length extending substantially through the test chamber diameter;
and a test instrument having an instrument housing including a
cartridge receptacle for supporting the test cartridge, the test
instrument further comprising sweeping means for sweeping the
agitator vane about the test chamber pivot point through the test
sample in substantially the full test chamber volume; detecting
means for detecting a reduction of sweeping movement of the
agitator vane; and timing means for timing a coagulation test time
elapsed from the commencement of sweeping of the agitator vane
until the detection of reduction of the sweeping movement.
2. The system of claim 1, wherein: the test cartridge is formed of
a cartridge housing enclosing each test chamber into which a test
sample is to be introduced and comprising a test chamber pivot
element; and the agitator vane comprises an agitator pivot element
engaging the test chamber pivot element allowing rotational motion
of the agitator vane about a pivot axis at the pivot point.
3. The system of claim 2, wherein the agitator vane comprises first
and second agitator vane leaflets extending in opposed directions
away from the pivot axis through the agitator vane length
presenting agitator leaflet sweeping faces bounded by vane leaflet
edges against the test sample disposed in the test chamber during
rotation of the agitator vane leaflets.
4. The system of claim 3, wherein the agitator vane leaflets are
symmetric and shaped in the form of propeller blades and present
symmetric, curved propeller sweeping faces against the test sample
disposed in the test chamber during rotation of the agitator vane
leaflets.
5. The system of claim 3, wherein the agitator vane is tubular and
the agitator vane leaflets are symmetric and present symmetric
convex curved leaflet sweeping faces against the test sample
disposed in the test chamber during rotation of the agitator vane
leaflets.
6. The system of claim 3, wherein the agitator vane leaflets are
symmetric and present symmetric substantially flat leaflet sweeping
faces against the test sample disposed in the test chamber during
rotation of the agitator vane leaflets.
7. The system of claim 6, wherein the agitator vane comprises an
elongated U-shaped channel extending between opposed channel ends
and an agitator pivot element between the opposed channel ends.
8. The system of claim 6, wherein the vane leaflet edges are
notched.
9. The system of claim 3, wherein the agitator vane comprises an
elongated U-shaped channel extending between opposed channel ends
and an agitator pivot element between the opposed channel ends.
10. The system of claim 3, wherein the vane leaflet edges are
notched.
11. The system of claim 3, wherein the agitator vane leaflets are
symmetric and present symmetric substantially convex leaflet
sweeping faces against the test sample disposed in the test chamber
during rotation of the agitator vane leaflets.
12. The system of claim 11, wherein the agitator vane comprises an
elongated tube extending between opposed tube ends and an agitator
pivot element between the opposed tube ends.
13. The system of claim 3, wherein the agitator vane leaflets are
symmetric and present symmetric concave curved leaflet sweeping
faces against the test sample disposed in the test chamber during
rotation of the agitator vane leaflets.
14. The system of claim 3, wherein the agitator vane leaflets are
symmetric and present symmetric leaflet sweeping faces having
notched leaflet edges against the test sample disposed in the test
chamber during rotation of the agitator vane leaflets.
15. The system of claim 3, wherein the agitator vane leaflets are
formed by a substantially rectangular frame surrounding a leaflet
window, whereby the agitator vane leaflets present leaflet sweeping
faces having at least one window against the test sample disposed
in the test chamber during rotation of the agitator vane
leaflets.
16. The system of claim 15, wherein the frame supports a mesh
disposed across the leaflet window.
17. The system of claim 3, wherein the agitator vane leaflets have
a plurality of sieve holes extending through the agitator vane
leaflets, whereby the agitator vane leaflets present leaflet
sweeping faces having a plurality of sieve holes against the test
sample disposed in the test chamber during rotation of the agitator
vane leaflets.
18. The system of claim 3, wherein the agitator vane leaflets have
a plurality of slots extending through the agitator vane leaflets,
whereby the agitator vane leaflets present leaflet sweeping faces
having a plurality of substantially parallel teeth against the test
sample disposed in the test chamber during rotation of the agitator
vane leaflets.
19. The system of claim 18, wherein the plurality of substantially
parallel teeth of each vane leaflet extend away from the pivot
axis.
20. The system of claim 3, wherein a mesh is disposed across the
agitator leaflet sweeping faces.
21. A test cartridge for use in a system that performs a test on a
test sample determining a coagulation test time comprising: a
cartridge housing having at least one cylindrical test chamber
therein into which a test sample is deposited to be tested, the
test chamber having a test chamber axis, a test chamber diameter
and a test chamber height defining a test chamber volume; and an
agitator mounted at a pivot point at the test chamber axis, the
agitator having an agitator vane adapted to be swept about the
pivot point and through the test sample in the test chamber, the
agitator vane having an agitator vane height extending
substantially through the test chamber height and an agitator vane
length extending substantially through the test chamber diameter,
whereby the agitator is adapted to be swept through substantially
the test chamber volume and contacts the test sample therein and a
coagulation test time from the commencement of sweeping of the
agitator vane until the detection of reduction of the sweeping
movement is determinable.
22. The test cartridge of claim 21, wherein: the test cartridge is
formed of a cartridge housing enclosing each test chamber into
which a test sample is to be introduced and providing a test
chamber pivot element; and the agitator vane comprises an agitator
pivot element engaging the test chamber pivot element allowing
rotational motion of the agitator vane about a pivot axis at the
pivot point.
23. The test cartridge of claim 22, wherein the agitator vane
comprises first and second agitator vane leaflets extending in
opposed directions away from the pivot axis through the agitator
vane length presenting agitator leaflet sweeping faces bounded by
vane leaflet edges against the test sample disposed in the test
chamber during rotation of the agitator vane leaflets.
24. The test cartridge of claim 23, wherein the agitator vane
leaflets are symmetric and shaped in the form of propeller blades
and present symmetric curved propeller sweeping faces against the
test sample disposed in the test chamber during rotation of the
agitator vane leaflets.
25. The test cartridge of claim 23, wherein the agitator vane is
tubular and the agitator vane leaflets are symmetric and present
symmetric convex curved leaflet sweeping faces against the test
sample disposed in the test chamber during rotation of the agitator
vane leaflets.
26. The test cartridge of claim 23, wherein the agitator vane
leaflets are symmetric and present symmetric substantially flat
leaflet sweeping faces against the test sample disposed in the test
chamber during rotation of the agitator vane leaflets.
27. The test cartridge of claim 26, wherein the agitator vane
comprises an elongated U-shaped channel extending between opposed
channel ends and an agitator pivot element between the opposed
channel ends.
28. The test cartridge of claim 26, wherein the vane leaflet edges
are notched.
29. The test cartridge of claim 23, wherein the agitator vane
comprises an elongated U-shaped channel extending between opposed
channel ends and an agitator pivot element between the opposed
channel ends.
30. The test cartridge of claim 23, wherein the vane leaflet edges
are notched.
31. The test cartridge of claim 23, wherein the agitator vane
leaflets are symmetric and present symmetric substantially convex
leaflet sweeping faces against the test sample disposed in the test
chamber during rotation of the agitator vane leaflets.
32. The test cartridge of claim 31, wherein the agitator vane
comprises an elongated tube extending between opposed tube ends and
an agitator pivot element between the opposed tube ends.
33. The test cartridge of claim 23, wherein the agitator vane
leaflets are symmetric and present symmetric concave curved leaflet
sweeping faces against the test sample disposed in the test chamber
during rotation of the agitator vane leaflets.
34. The test cartridge of claim 23, wherein the agitator vane
leaflets are symmetric and present symmetric leaflet sweeping faces
having notched leaflet edges against the test sample disposed in
the test chamber during rotation of the agitator vane leaflets.
35. The test cartridge of claim 23, wherein the agitator vane
leaflets are formed by a substantially prismatic frame surrounding
a leaflet window, whereby the agitator vane leaflets present
leaflet sweeping faces having at least one window against the test
sample disposed in the test chamber during rotation of the agitator
vane leaflets.
36. The test cartridge of claim 35, wherein the frame supports a
mesh disposed across the leaflet window.
37. The test cartridge of claim 23, wherein the agitator vane
leaflets have a plurality of sieve holes extending through the
agitator vane leaflets, whereby the agitator vane leaflets present
leaflet sweeping faces having a plurality of sieve holes against
the test sample disposed in the test chamber during rotation of the
agitator vane leaflets.
38. The test cartridge of claim 23, wherein the agitator vane
leaflets have a plurality of slots extending through the agitator
vane leaflets, whereby the agitator vane leaflets present leaflet
sweeping faces having a plurality of substantially parallel teeth
against the test sample disposed in the test chamber during
rotation of the agitator vane leaflets.
39. The test cartridge of claim 38, wherein the plurality of
substantially parallel teeth of each vane leaflet extend away from
the pivot axis.
40. The test cartridge of claim 23, wherein a mesh is disposed
across the agitator leaflet sweeping faces.
41. A method of performing a test on a test sample and determining
a coagulation test time comprising: providing a test cartridge
comprising: a cartridge housing having at least one cylindrical
test chamber therein into which a test sample is deposited to be
tested, the test chamber having a test chamber axis, a test chamber
diameter and test chamber height defining a test chamber volume;
and an agitator mounted at a pivot point at the test chamber axis,
the agitator having an agitator vane adapted to be swept about the
pivot point and through the test sample in the test chamber unless
impeded by coagulation of the test sample, the agitator vane having
an agitator vane height extending substantially through the test
chamber height and an agitator vane length extending substantially
through the test chamber diameter; and filling the test chamber
with a test sample; inserting the test cartridge into a cartridge
receptacle of a test instrument; and operating the test instrument
to: sweep the agitator vane about the test chamber pivot point
through the test sample; detect the reduction of sweeping movement
of the agitator vane; and time a coagulation test time elapsed from
the commencement of sweeping of the agitator vane until the
detection of reduction of the sweeping movement.
42. A method of performing a test on a biologic test sample and
determining a coagulation test time comprising: providing a test
cartridge comprising: a cartridge housing having at least one
cylindrical test chamber therein into which a test sample is
deposited to be tested, the test chamber having a test chamber
axis, a test chamber diameter and test chamber height defining a
test chamber volume; and an agitator mounted at a pivot point at
the test chamber axis, the agitator having an agitator vane adapted
to be swept about the pivot point and through the test sample in
the test chamber unless impeded by coagulation of the test sample,
the agitator vane having an agitator vane height extending
substantially through the test chamber height and an agitator vane
length extending substantially through the test chamber diameter;
and filling the test chamber with a test sample; inserting the test
cartridge into a cartridge receptacle of a test instrument; and
operating the test instrument to: sweep the agitator vane about the
test chamber pivot point through the test sample; detect the
sweeping movement of the agitator vane; and display an image of the
sweeping movement of the agitator vane, whereby changes in the
sweeping movement due to coagulation and dissolution of coagulation
are presented.
43. The method of claim 42, wherein the test sample is a blood test
sample and the dissolution of coagulation is due to fibrinolysis of
coagulation.
Description
REFERENCE TO RELATED APPLICATION
[0001] Reference is hereby made to commonly assigned U.S.
Provisional Application No. (P-7928.00) filed on Apr. 19, 2004 for
BLOOD COAGULATION TEST CARTIDGE, SYSTEM, AND METHOD in the name of
David Wright et al.
FIELD OF THE INVENTION
[0002] This invention relates to detecting changes in viscosity of
biologic fluid test samples, e.g., detecting coagulation and
coagulation-related activities including agglutination and
fibrinolysis of human blood test samples, and more particularly to
improved methods and apparatus for obtaining a coagulation time of
a blood test sample.
BACKGROUND OF THE INVENTION
[0003] Blood coagulation is a complex chemical and physical
reaction that occurs when blood (herein, "blood" shall mean whole
blood, citrated blood, platelet concentrate, plasma, or control
mixtures of plasma and blood cells, unless otherwise specifically
called out) comes into contact with an activating agent, such as an
activating surface or an activating agent. In accordance with one
simplified conceptual view, the whole blood coagulation process can
be generally viewed as three activities: platelet adhesion,
platelet aggregation, and formation of a fibrin clot. In vivo,
platelets flow through the blood vessels in an inactivated state
because the blood vessel lining, the endothelium, prevents
activation of platelets. When a blood vessel is damaged, however,
the endothelium loses its integrity and platelets are activated by
contact with tissue underlying the damaged site. Activation of the
platelets causes them to become "sticky" and adhere together.
Additional platelets then adhere to the activated platelets and
also become activated. This process continues until a platelet
"plug" is formed. This platelet plug then serves as a matrix upon
which blood clotting or coagulation proceeds. Blood "coagulation"
and "clotting" are used interchangeably herein unless specifically
distinguished from one another.
[0004] If the chemical balance of the blood is suitable, thrombin
is then produced that causes fibrinogen to convert to fibrin, which
forms the major portion of the clot mass. During clotting,
additional platelets are activated and trapped in the forming clot,
contributing to clot formation. As clotting proceeds,
polymerization and cross-linking of fibrin results in the permanent
clot. Thus, platelet activation plays a very important function in
blood coagulation.
[0005] The clinical assessment of clotting function has long been
recognized to be important in the management of surgical patients.
Preoperatively, the assessment of the clotting function of the
patient's blood is utilized as a predictor of risk of patient
bleeding, allowing advanced preparation of blood components.
Perioperative monitoring of the clotting function of the patient's
blood is also important because coagulopathies can be induced by
hemodilution of procoagulants, fibrinogen and platelets, by
consumption of coagulation factors during surgical procedures, or
by cardiopulmonary bypass. Post-operative assessment of clotting
function is also crucial to the patient's successful recovery. For
example, 3-5% of cardiopulmonary bypass patients require surgical
reoperation to stop bleeding. Prompt assessment of clotting
function could rule out coagulopathy as the cause of bleeding and
could avoid unnecessary surgery that adds to patient morbidity and
treatment costs.
[0006] Several tests of coagulation are routinely utilized to
assess the complicated cascade of events leading to blood clot
formation and test for the presence of abnormalities or inhibitors
of this process. Among these tests are platelet count (PLT),
thrombin time (TT), prothrombin time (PT), partial thromboplastin
time (aPTT), activated clotting time (ACT), fibrinogen level (FIB),
fibrinogen degradation product concentrations, and general purpose
clotting (GPC) time. The aPTT test can be used to assess the degree
of anticoagulation resulting from heparin administration. The PT
test results can indicate the level of anticoagulation produced by
warfarin (coumadin) administration. In the GPC test, the blood
sample is exposed to calcium as an activating agent, but other
reagents can be added to conduct the aPTT test, for example.
[0007] The ACT tests are further differentiated into high range ACT
(HRACT), low rate ACT (LRACT), recalcified ACT (RACT), and HRHTC
tests. The HRACT test features a slope response to moderate to high
heparin levels in whole blood drawn from a patient during cardiac
surgery for use in whole blood coagulation time tests. The LRACT
test features a more sensitive slope response to heparin levels
ideal for use in whole blood coagulation time tests run in the ICU,
CCU and during dialysis. The RACT test is similar to the LRACT
test, but the test is conducted on a citrated blood test sample,
rather than a whole blood sample, and is typically conducted at a
central testing station remote from the patient the test sample is
drawn from.
[0008] The HRHTC test is a variation upon the HRACT test where an
enzyme, heparinase, is added as a deactivating or neutralizing
reagent to one test sample of the whole blood drawn from the
patient to neutralize any heparin therein and thereby provide a
heparin-free blood test sample. In this test, comparative
coagulation times are determined from the heparinase-exposed blood
test sample and the unexposed blood test sample. The two ACT test
times will be close if no heparin is present in the whole blood
drawn from the patient. The ACT test conducted on the
heparinase-exposed blood test sample will result in a shorter
coagulation test time than the ACT test conducted on the unexposed
blood test sample, thereby establishing that the patient's blood
contains at least some level of heparin.
[0009] During heart bypass surgery, the platelets of blood
circulated in an extracorporeal circuit may become activated by
contact with the materials present in the extracorporeal circuit.
This activation may be reversible or irreversible. Once platelets
are irreversibly activated, they lose their ability to function
further. A deficiency of functional platelets in the blood may be
indicative of an increased probability of a post-operative bleeding
problem. Such a deficiency, and the resulting post-operative
bleeding risk, could be remedied by a transfusion of platelet
concentrate. Platelet functionality tests, e.g., the ACT test, can
identify a deficiency of platelets or functional platelets and aid
the attending surgeon in ascertaining when to administer a platelet
concentrate transfusion. Such a test is further useful in
ascertaining the efficacy of a platelet transfusion. By performing
the platelet functionality test following a platelet transfusion,
it is possible to determine if additional platelet concentrate
transfusions are indicated. Real-time assessment of clotting
function at the operative site is preferred to evaluate the result
of therapeutic interventions and also to test and optimize, a
priori, the treatment choice and dosage.
[0010] A number of different medical apparatus and testing methods
have been developed for measuring and determining platelet
activation and coagulation-related conditions of blood that can be
used in real time during surgery, particularly bypass surgery, on
fresh drawn blood samples or that can be used after some delay on
citrated blood samples. Some of the more successful techniques of
evaluating blood clotting and coagulation of fresh or citrated
blood samples employ plunger techniques disclosed in commonly
assigned U.S. Pat. Nos. 4,599,219, 4,752,449, 5,174,961, 5,314,826,
5,925,319, and 6,232,127, for example. These techniques are
embodied in the ACT II.RTM. automatic coagulation timer,
commercially sold by the assignee of this patent application.
[0011] In U.S. Pat. No. 5,302,348, an apparatus and method are
disclosed for performing a coagulation time test on a sample of
blood deposited in a fluid reservoir of a disposable cuvette. A
capillary conduit having at least one restricted region is formed
within the cuvette. The cuvette is inserted into a testing machine
that engages the cuvette and draws blood from the fluid reservoir
into the capillary conduit. The blood is then caused to
reciprocally move back and forth within the capillary conduit so
that the blood is forced to traverse the restricted region. Optical
sensors of the testing machine are employed to detect movement of
the blood. The testing machine measures the time required each time
the blood is caused to traverse the restricted region. Coagulation
is considered to have occurred and the overall coagulation time is
displayed to the operator when a measured time is a predetermined
percentage longer than an immediately preceding time.
[0012] In U.S. Pat. No. 5,504,011, a similar apparatus and method
are disclosed for performing multiple coagulation time tests on a
sample of blood deposited in a fluid reservoir of a disposable
cuvette having multiple capillary conduits within the cuvette. Each
of the conduits contains a dried or lyophilized activation reagent
that is rehydrated by the blood. The blood in each conduit is then
reciprocally moved across a restricted region of the conduit until
a predetermined degree of coagulation occurs. Since the coagulation
time is being monitored in multiple conduits, a representation
coagulation time for a given sample can be determined. A
normalizing control agent is present in at least one of the
conduits. The normalizing control agent counteracts any effects of
anticoagulants present in the blood sample, thereby allowing the
blood sample to have generally normal coagulation characteristics.
The normalized blood is tested simultaneously with the untreated
blood to provide a reference value against which the functionality
of the test system and the quality of the sample can be judged.
[0013] The apparatus and methods disclosed in the '348 and '011
patents only check the state of the sample during the reciprocal
back and forth movement of the sample through the restricted region
capillary. The detection of coagulation would be delayed or
inaccurate if the sample coagulates between movement cycles.
[0014] In U.S. Pat. No. 6,200,532, a device and method for
performing blood coagulation assays, particularly prothrombin times
and activated partial thromboplastin times and other clotting
parameters are disclosed. One embodiment of the device comprises a
disposable cassette containing a sample inlet for sample delivery,
a pair of interleaved spiral capillary channels for driving force,
and a reaction chamber with an appropriate dry reagent for a
specific assay, and a piezoelectric sensor. The device could also
include a heating element for temperature control, and a magnetic
bender. Compressed air is employed to drive the sample into the two
spiral capillary channels. The magnetic bender is driven by an
electromagnetic field generator and is attached onto a
piezoelectric film in contact with the blood sample. The electric
signal generated in the piezoelectric film is characterized by its
frequency and amplitude due to the movement of the attached metal
film. The signal collected at the site of the piezoelectric film
represents the process of a biochemical reaction in the reaction
chamber as the blood sample proceeds to the point at which clot
formation starts and is amplified by an amplifier and rectified
into a DC voltage and is sent to a recording unit and/or display
unit.
[0015] A method and apparatus for measuring coagulation of blood is
disclosed in U.S. Pat. No. 4,879,432, wherein a timer is started as
a stream of solid particles are introduced into a tube containing
the blood sample that descend through the blood sample under the
force of gravity. The particles are micron-sized grains of glass,
for example, that are highly wettable and can descend through the
blood sample until stopped by the fibrin network that forms as
coagulation begins. Photoelectric cells detect the stopping of the
particles, and the coagulation time is determined.
[0016] Other approaches to the detection of a change in viscosity
of a fluid, particularly changes accompanying fibrin formation and
clotting in blood samples, have been disclosed involving
ferromagnetic elements or materials introduced in the fluid that
are inhibited from moving freely when the viscosity change or
clotting occurs.
[0017] For example, a method and apparatus is disclosed in U.S.
Pat. No. Re. 27, 866 wherein the coagulation of a blood plasma
sample is intensified by dispersing iron oxide particles throughout
the plasma sample. A rotating magnetic field is applied that causes
the particles to move within the sample and activate the clotting
reaction. The moving particles collect fibrin strands that are
formed, thereby changing the optical properties of the
particle-sample mixture, and the changed optical properties are
detected optically.
[0018] Further methods and apparatus employing magnetic effects for
determining changes in viscosity of a fluid, particularly a blood
sample, within a test tube are disclosed in U.S. Pat. Nos.
3,635,678, 3,836,333, 3,967,934, and 5,145,082. In the '678 and
'934 patents, a steel ball within the fluid is maintained at a
certain location in the test tube by a magnetic field, while the
tube is moved up and down. The magnetic force is insufficient to
attract the ball when the fluid in the tube changes viscosity, and
the failure of the steel ball to stay within the magnetic field is
detected photoelectrically. In the '333 and '082 patents, a blood
sample is placed in a test tube along with a ferromagnetic member.
A magnetic field is applied to the ferromagnetic member, and the
applied magnetic field maintains the ferromagnetic member oriented
with the magnetic field as the test tube is rotated about its axis.
The magnetic force is insufficient to attract the ball when the
fluid in the tube changes viscosity, e.g., due to clotting, and the
failure of the member to stay within the magnetic field is detected
magnetically with a reed switch.
[0019] In other approaches to measuring coagulation time, a blood
sample is disposed in a test chamber having a ferromagnetic element
therein. The ferromagnetic element is raised against gravity, and
the field is discontinued so that the ferromagnetic member drops
through the sample in the test chamber.
[0020] For example, a viscometer is disclosed in U.S. Pat. No.
4,648,262 wherein the fluid sample under test is placed within a
capillary tube along with a metal ball. A magnet mounted on a
rotating drum periodically raises the ball to the top of the tube
and then releases the ball so that it can fall to the bottom of the
tube. The viscosity of the fluid is determined by the rate of
descent of the ball in the tube.
[0021] Apparatus and methods for detecting changes in human blood
viscosity are disclosed in U.S. Pat. Nos. 5,629,209 and 6,613,286,
wherein heparinized blood is introduced into a test cartridge
through an injection port and fills a blood receiving/dispensing
reservoir. The blood then moves from the reservoir through at least
one conduit into at least one blood-receiving chamber where it is
subjected to a viscosity test. A freely movable ferromagnetic
washer is also located within the blood-receiving chamber that is
moved up using an electromagnet of the test apparatus and allowed
to drop with the force of gravity. Changes in the viscosity of the
blood that the ferromagnetic washer falls through are detected by
determining the position of the ferromagnetic washer in the
blood-receiving chamber over a given time period or a given number
of rises and falls of the ferromagnetic washer. The blood sample
can be mixed with a viscosity-altering agent (e.g., protamine) as
it passes through the conduit to the blood-receiving chamber. Air
in the conduit and blood-receiving chamber is vented to atmosphere
through a further vent conduit and an air vent/fluid plug as the
blood sample is fills the blood-receiving chamber.
[0022] The movement of the washer in the above approach is actively
controlled only when it is moved up, and the washer passively drops
with the force of gravity. The washer is free to float in the test
chamber and may drift side-to-side as it is moved up or floats
downward. The side-to-side drifting movement may affect the rise
time and the fall time, which could add error to the coagulation
time measured. The washer may eventually stop moving as a clot
forms about it, and no additional information can be obtained on
the coagulation process in the sample.
[0023] It would be desirable to provide inexpensive, relatively
simple, easy to use, and accurate equipment and techniques that
quickly measure one or more of the aforementioned blood coagulation
times, including TT, PT, aPTT, and ACT.
SUMMARY OF THE INVENTION
[0024] Therefore, the present invention provides a compact,
inexpensive, disposable, test cartridge in which a test sample,
particularly a blood or biologic test sample, to be tested is
deposited and systems and methods for detecting a change in test
sample viscosity. In a particular application, the present
invention provides systems and methods for measuring a test
coagulation time of such a test sample that are simple to practice,
rapid, and reliable.
[0025] The test sample may comprise a biologic test sample or a
blood test sample, e.g., whole blood drawn from a patient or plasma
or blood concentrate or citrated blood, and the blood test sample
can be maintained at body temperature, can be heparinized, and can
be mixed with a reagent. The reagent is selected from coagulants or
activating agents including calcium, kaolin, celite, ellagic acid,
glass particles, thrombin, thromboplastin or other activating
agents or from heparin neutralizing or deactivating agents
including heparinase or protamine. The test coagulation time may
comprise one or more of the aforementioned blood coagulation times,
including TT, PT, aPTT, and ACT.
[0026] The test cartridge comprises a cartridge housing having at
least one test chamber therein into which a test sample is
deposited to be tested. An agitator vane of an agitator is mounted
at a pivot point and is adapted to be swept about the pivot point
and through the test sample in the test chamber when the test
sample is not coagulated. A coagulation test time from the
commencement of sweeping of the agitator vane until the detection
of cessation or reduction of the sweeping movement is
determinable.
[0027] In determining a coagulation time, a timer is started after
a test sample is deposited in the test chamber and the agitator
vane movement is commenced. The sweeping movement mixes the test
sample, and the agitator vane initially sweeps through the test
sample without encountering significant resistance. Resistance to
agitator vane movement signifies coagulation of the test sample.
The elapsed time measured from starting the sweeping movement and
the detection of resistance constitutes a coagulation time.
[0028] The resistance to movement leads to a change or reduction in
agitator vane movement, e.g., slowing agitator vane movement,
reducing the range of the agitator vane movement or halting
agitator vane movement. The reduction in agitator vane movement,
including slowing of sweeping movement, reduction in range of
movement, and absence or cessation of sweeping movement, is
preferably detected by one or more agitator vane sensor and
agitator movement detection circuitry coupled thereto. The agitator
vane sensor may comprise one of a proximity sensor, inductive
sensor, magnetic sensor, metal sensor, and optical sensor disposed
at a sensor location with respect to the test chamber. Optionally,
the detected movement of the agitator vane is displayed for
observation of changes in agitator vane movement over time.
[0029] In a preferred embodiment, the test cartridge is formed of a
cartridge housing having a substantially prismatic shape with upper
and lower major sides and a minor sidewall extending between the
upper and lower major sides and maintaining the upper and lower
major sides in substantially parallel relation. Each test chamber
extends between the upper and lower major sides in a portion of the
cartridge housing thereby having a test chamber height. A test
sample is deposited in a cartridge port and travels through a
filling channel or capillary into the test chamber. Preferably, an
air vent channel or capillary extends from the test chamber to an
air vent.
[0030] In preferred embodiments, the agitator has an agitator pivot
element engaging a test chamber pivot element defining the pivot
point and a pivot axis, whereby the agitator vane extends from the
agitator pivot element intermediate the upper and lower major sides
and can be swept about the pivot axis at the pivot point in the
absence of coagulation.
[0031] The agitator vane preferably comprises first and second
agitator vane leaflets extending in opposed directions away from
the agitator pivot element, the vane leaflets having a leaflet
width, a leaflet thickness, and a leaflet length. The leaflet
length is somewhat smaller than the test chamber radius, whereby
the agitator vane leaflet extends from the agitator pivot element
substantially across the test chamber radius. The vane leaflet
height is somewhat smaller than the test chamber height, and the
leaflet thickness is less than the vane leaflet height and the
leaflet length. Thus, the preferred agitator vane shape possesses
agitator leaflets that present enlarged agitator leaflet sweeping
faces against the test sample disposed in the test chamber during
rotation of the agitator vane leaflets.
[0032] The agitator vane leaflets have leaflet faces extending
across the leaflet width and length that bear against and sweep
through the fluid test sample as the agitator vane leaflets are
swept about the pivot point within the test chamber. The leaflet
faces thereby present high fluid interface profiles to the fluid
test sample as the fluid test sample washes against the vane
leaflet faces during the sweeping movement of the agitator vane
leaflets about the pivot point and through the test sample.
[0033] In certain preferred embodiments, the agitator vane leaflets
preferably are shaped as paddles or blades that have first and
second leaflet faces that are substantially parallel to one another
and are substantially parallel to the pivot axis. The leaflet faces
can be solid and impermeable by the test sample or can be permeable
to the test sample. The agitator vane leaflets can have leaflet
edges that are relatively smooth or are notched or roughened.
[0034] In a further preferred embodiment, the agitator vane is
preferably formed shaped like a propeller wherein the vane leaflets
comprise propeller blades each having a blade length, blade width
and a blade thickness, and the propeller blades present blade faces
that are obliquely angled to the pivot axis.
[0035] In still further preferred embodiments, the agitator vane
leaflets are each preferably a U-shaped channel extending away from
the agitator pivot element.
[0036] In yet another preferred embodiment, the agitator vane
leaflets are each preferably tubular extending away from the
agitator pivot element.
[0037] The agitator vane is preferably ferromagnetic, and the
sweeping movement of the agitator vane about the pivot point is
preferably effected by a sweeping magnetic field that captures and
sweeps a ferromagnetic agitator vane leaflet about the pivot point.
The sweeping magnetic field is preferably formed by a plurality of
electromagnets disposed in a test instrument in relation to a test
cartridge receptacle shaped to receive the test cartridge.
Electrical sweeping signals are applied to the electromagnets to
generate the sweeping magnetic field that effects the sweeping
movement. The magnetic field strength is sufficient to impart
sweeping movement of the agitator vane when resistance is low in
the absence of coagulation of the test sample and is insufficient
to overcome resistance to the agitator vane movement that develops
with coagulation of the test sample.
[0038] During a coagulation time test, a sweeping magnetic field
can be generated that effects continuous rotation of the agitator
vane through 360.degree. revolutions at a predetermined revolution
per minute about the pivot point (when the pivot point is disposed
in the center of a cylindrical test chamber). In one preferred
method, the sweeping movement comprises alternating clockwise and
counter-clockwise sweeps of the agitator vane from a neutral
position through predetermined arcs of less than 360.degree. in
response to a sweeping signal applied to the electromagnet(s) at a
predetermined sweep rate.
[0039] In either case, at least one agitator vane sensor is
disposed in operative relation to at least one agitator vane and
provides an agitator sensor output signal upon each sweep of the
agitator vane past the agitator sensor. The agitator sensor output
signal waveform varies in signal parameters, e.g., signal amplitude
and cycle length, correlated in time to the applied sweeping signal
during the coagulation time test. Preferably, the output signal
waveform is sampled, digitized and stored as a reference or
template waveform during a template establishing time period.
Subsequently generated agitator sensor output signals are
continually sampled, digitized, and compared to the template
waveform. Coagulation is declared when the waveform fails to
substantially match the template waveform while the sweeping signal
continues to be applied to the electromagnets. A test failure or
maximum clotting time is declared if the waveform of the
subsequently generated agitator sensor output signal continues to
substantially match the template waveform over a predetermined
maximum time window following commencement of the coagulation time
test.
[0040] In a preferred embodiment combining both vane movements and
wherein the test chamber is cylindrical, a sweeping signal is
generated that effects continuous rotation of the agitator vane
through 360.degree. at a predetermined revolution per minute in an
initial mixing phase in order to mix the test sample with a
reagent, if present. A test phase commences upon completion of the
initial mixing phase, wherein the sweeping movement comprises
alternating clockwise and counter-clockwise sweeps of the agitator
vane from a neutral position through predetermined arcs of less
than 360.degree. in response to a sweeping signal applied to the
electromagnets at a predetermined sweep rate. The coagulation time
is measured either from the commencement of the mixing phase or the
test phase until the resistance to sweeping movement is detected. A
test failure is declared if the amplitude of the sensor output
signal does not vary during the initial mixing phase, and no
template waveform can be determined.
[0041] In preferred embodiments, at least two test chambers are
included in each test cartridge, and redundant tests are performed
simultaneously in a test instrument to provide increased accuracy.
Other embodiments of the test cartridge may include additional test
chambers to increase redundancy or to perform differing tests on
the same test sample simultaneously or sequentially.
[0042] The test sample may be first mixed with the reagent before
deposit in the test chamber, or may contact the reagent coating on
the surfaces of an filling channel extending between the blood
receptacle and the test chamber. Additionally or alternatively, the
test sample may be mixed with a reagent disposed in the test
chamber. For example, the agitator vane or surfaces of the test
chamber may be coated with the reagent. Or, a discrete element that
is coated with or comprises a reagent is disposed in the test
chamber. A plurality of test cartridges can be supplied with
particular reagents and identified by color or indicia for
conducting specific tests of test samples. In a multi-chamber test
cartridge, certain of the test chambers may be devoid of any
reagent and other test chambers may include a reagent or differing
concentrations of reagents.
[0043] Moreover, the agitator vane can be surface etched to
increase surface area and to enhance surface roughness that tends
to cause fibrin to develop in blood test samples. The surface
roughness may activate the coagulation of the sample in place of or
in addition to a reagent.
[0044] Advantageously, the agitator vane is actively swept through
the test sample in response to the generated magnetic fields and
does not depend on the force of gravity to passively move the
agitator vane through the test sample. The continuous, active
control of movement of the agitator vane reduces errors introduced
with passive control in prior art systems that rely upon
gravity.
[0045] In addition, the active driving force may be applied to
induce the agitator vane to rotate again after the reduction or
cessation of movement of the agitator vane is detected to track the
continued formation of the clot and the effect of the fibrinolytic
process (the breakdown of the clot). The agitator vane may begin to
move again due to the applied active driving force as fibrinolysis
occurs to weaken the coagulation forces resisting agitator vane
movement. The mechanical integrity of the clot over a longer period
of time can be assessed, particularly to identify coagulopathic
characteristics of the blood test sample.
[0046] The agitator vane sensor continuously monitors agitator vane
movement thereby capturing any change in movement as soon as the
change occurs. In this way, there is no lag time between a change
in agitator vane movement and detection of the change.
[0047] The present invention is useful for providing information
concerning the discrete events that may accompany the coagulation,
clotting or lytic process (e.g., clot formation, clot retraction,
or clot lysis), as well as provide insight into the overall event.
The present invention may be useful in distinguishing between
platelet-rich and platelet-poor plasma depending on the clotting
profile of the sample. The present invention may also provide
information useful to physicians developing a prognosis for
patients following cardiopulmonary bypass surgery to avoid or
mitigate post-operative bleeding.
[0048] This summary of the invention has been presented here simply
to point out some of the ways that the invention overcomes
difficulties presented in the prior art and to distinguish the
invention from the prior art and is not intended to operate in any
manner as a limitation on the interpretation of claims that are
presented initially in the patent application and that are
ultimately granted.
BRIEF DESCRIPTION OF THE DRAWINGS
[0049] These and other advantages and features of the present
invention will be more readily understood from the following
detailed description of the preferred embodiments thereof, when
considered in conjunction with the drawings, in which like
reference numerals indicate identical structures throughout the
several views, and wherein:
[0050] FIG. 1 is a perspective view of a test instrument for
receiving a test cartridge and for performing tests on test samples
deposited in test chambers of the test cartridge, particularly to
determine a blood coagulation time;
[0051] FIG. 2 is a perspective view of one embodiment of a
prismatic blood coagulation test cartridge of the present invention
having two test chambers;
[0052] FIG. 3 is an exploded perspective view of the components of
the test cartridge of FIG. 2 including the agitators fitted into
the test chambers;
[0053] FIG. 4 is an exploded side view of the components of the
test cartridge of FIGS. 2 and 3;
[0054] FIG. 5 is a side cross-section view taken along lines 5-5 of
FIG. 4;
[0055] FIG. 6 is a side cross-section view taken along lines 6-6 of
FIG. 2;
[0056] FIG. 7 is an enlarged perspective view of the agitators
employed in the test cartridge of FIGS. 2-6;
[0057] FIGS. 8-12 are top views of an agitator employed in the test
cartridge of FIGS. 1-6 schematically depicting electromagnetic
field poles energized by phased sweeping signals for rotating the
agitator vane leaflets about the pivot point and the location of an
agitator vane sensor disposed to detect to the motion of the
agitator vane leaflets;
[0058] FIG. 13 is a diagram of agitator sensor output signals
observed in a coagulation time test employing the test cartridge of
FIGS. 1-6 during rotation as depicted in FIGS. 8-12 and upon
cessation of rotation;
[0059] FIG. 14 is a flowchart illustrating the steps of operation
of the system of FIG. 1 employing a test cartridge of FIGS.
1-6;
[0060] FIG. 15 is a top view of a second embodiment of an agitator
employed in a test cartridge of the type depicted in FIGS. 1-6 or
FIG. 16 adapted to be swept about the pivot point in relation to an
agitator vane sensor as in FIGS. 8-12 by schematically depicted
electromagnetic field poles energized by phased sweeping signals
that rotate the agitator vane leaflets, wherein the agitator
leaflets present enlarged agitator leaflet sweeping faces against
the test sample disposed in the test chamber during rotation of the
agitator vane leaflets;
[0061] FIG. 16 is a cross-section view of the second embodiment of
the agitator in relation to the test chamber of FIG. 15;
[0062] FIG. 17 is an exploded view of a modified test cartridge of
the present invention employing a first variation of the second
embodiment of the agitator;
[0063] FIGS. 18-26 are perspective views of alternative preferred
agitator embodiments incorporating dimensional features of the
agitator of FIG. 16 usable in the test cartridge embodiment of
FIGS. 1-6 or FIG. 16; and
[0064] FIG. 27 is a diagram of agitator sensor output signals
observed in a coagulation time test showing the effects of
coagulation and subsequent fibrinolysis upon movement of an
agitator vane through the test sample.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE
INVENTION
[0065] In the following detailed description, references are made
to illustrative embodiments of methods and apparatus for carrying
out the invention. It is understood that other embodiments can be
utilized without departing from the scope of the invention.
Preferred methods and apparatus are described for performing blood
coagulation tests of the type described above.
[0066] FIG. 1 is perspective view of the simplified automatic
coagulation time test instrument 10 that can advantageously be used
in the practice of the present invention. The test instrument 10 is
portable and is operated by battery power or alternatively by power
from an AC line. The test instrument 10 has a cartridge receptacle
24 that receives a test cartridge 50 shown in more detail FIGS. 2-6
inserted through the external slot of he cartridge receptacle 24.
The test instrument 10 can incubate test samples within the test
chambers of the test cartridge 50. The test instrument 10 also
conducts coagulation time tests on test samples injected from the
same source into two test chambers of the test cartridge 50. The
test samples are subjected to the same reagents for conducting
redundant coagulation time tests or differing reagents for
conducting comparative coagulation time tests. The parallel tests
are designated "Channel 1" and "Channel 2" coagulation time
tests.
[0067] For consistency and accuracy, certain coagulation time tests
are conducted on test samples that are "incubated" in the test
chambers of the test cartridge 50 by maintaining the test samples
at body temperature for a defined incubation time period. It may be
necessary to incubate citrated whole blood, plasma, or quality
control samples prior to running ACT tests because they may be
chilled or at room temperature. Heat is provided also when
conducting ACT tests of fresh whole blood employing the test
instrument 10 so that the ACT tests are consistently performed at
37.degree. C. (body temperature) rather than at blood test sample
temperatures that are elevated or depressed from 37.degree. C.
Blood test samples withdrawn from a patient and immediately
injected into the test cartridge before the sample has cooled may
not need an incubation period and may only need to be maintained at
37.degree. C.
[0068] Thus, a heating block (not shown) is incorporated in the
test instrument 10 in relation to the cartridge receptacle 24.
Preferably the heating block is continuously powered when the test
instrument 10 is powered in order to maintain a constant
temperature, which is nominally body temperature or 37.degree. C.,
to a test cartridge 50 inserted into the cartridge receptacle 24.
The heating block may be mounted to be manually moved into
operative relation with the cartridge receptacle 24 to apply heat
to a test cartridge 50 inserted into operative heating relation to
the cartridge receptacle 24 or away from cartridge receptacle 24 in
the manner of the heating block of the MEDTRONIC.RTM. ACT II
automated coagulation timer sold by Medtronic, Inc. An Incubate
switch 16 is located on the front panel 14 that can be depressed to
time out incubation for a predetermined incubation time. An amber,
backlit, indicator designated "On" is illuminated when the Incubate
switch 16 is activated, and the numerical temperature display 28
displays the temperature of the heating block, which is normally
37.degree. C., incubation temperature.
[0069] The front panel 14 also supports a Mix switch 20 and a Start
switch 22. The clinician activates the Mix switch 20 when an
initial mixing phase is to be included in the coagulation time
test. A further amber, backlit, indicator designated "On" is
illuminated when the Mix switch 20 is activated. The Start switch
22 is depressed by the clinician to commence a coagulation time
test with or without a mixing phase. Additional switches and
indicators may be provided on front panel 14 to conduct additional
tests or additional analysis beyond determination of the
coagulation time of the test sample or test samples.
[0070] An LCD screen 26 and four amber, backlit indicators
positioned alongside upper and lower display lines of the LCD
screen 26 are also supported by the front panel 14. The upper and
lower display lines of LCD screen 26 display Channel 1 and Channel
2 incubation or coagulation times. The elapsed or remaining
incubation times are displayed for the Channel 1 and Channel 2 test
samples when incubation is taking place. The elapsed Channel 1 and
Channel 2 coagulation test times are displayed for the Channel 1
and Channel 2 test samples as the coagulation times are being timed
out. The final Channel 1 and Channel 2 coagulation test times are
then displayed for the Channel 1 and Channel 2 test samples when
coagulation is detected.
[0071] In addition, the "Average" and "Difference" values of the
final Channel 1 and Channel 2 coagulation test times are displayed
as the Display Switch 22 is successively depressed. The "Average"
and "Difference" can only be displayed by depressing push button
display switch 22 upon determination and display of the separate
final "Channel 1" and "Channel 2" coagulation times for the two
test samples. In each case, the corresponding indicators are
backlit to identify the displayed data.
[0072] It should be noted that the test instrument operating
algorithm could also calculate and display the percent difference
between the Channel 1 and Channel 2 coagulation times on the LCD
screen 26. A percentage difference greater than a certain threshold
(12% in the case of the ACT test) indicates that the coagulation
time measurements are "out of tolerance" and that the test should
be repeated with a new test sample.
[0073] In addition, the upper and lower display lines of LCD screen
26 display Channel 1 and Channel 2 test failure alarms if the
coagulation times are too short. For example, a test failure is
declared if a coagulation sensor detects coagulation during an
initial mixing phase or otherwise too early in the testing phase of
one or both of the Channel 1 and Channel 2 coagulation tests.
[0074] The front panel 14 also supports a Stop switch 18 that can
be depressed by the clinician to terminate either of the incubation
phase or the coagulation time test that is in progress. The
incubation or coagulation test times that are displayed in display
26 when the Stop switch 18 is depressed are frozen and the
displayed times flash.
[0075] The elapsed coagulation test times between 6 and 100 seconds
are displayed in 1/10 second resolution if the coagulation tests
are conducted following incubation of the test samples in the test
chambers of the test cartridge 20. Any coagulation test times
exceeding 100 seconds are displayed in whole seconds because the
1/10 resolution after 100 seconds is not deemed critical. The 1/10
second resolution is important if a PT test is being performed
because PT test times are typically short.
[0076] The sensors and circuitry of the test instrument 10 conduct
the coagulation time tests on the blood samples in the test
chambers of the two test chambers of the test cartridge 50 inserted
into the cartridge receptacle 24 in a manner described further
below.
[0077] A first embodiment of a test cartridge 50 used with the test
instrument depicted in FIG. 1 in accordance with the method of FIG.
14 is depicted in FIGS. 2-9. The test cartridge 50 is formed of a
cartridge housing 52 and other components described further below.
The cartridge housing 52 has a substantially prismatic shape with
upper and lower major sides 54 and 56 and a minor cartridge edge or
sidewall 58 extending between the upper and lower major sides 54
and 56 and maintaining the upper and lower major sides 54 and 56 in
substantially parallel relation. Finger grip indentations are
formed in the minor cartridge sidewall 58 to facilitate holding the
test cartridge 50 upright, filling test chambers with the test
sample, inserting and removing the cartridge housing 52 into and
from the instrument cartridge receptacle 24.
[0078] The cartridge housing 52 is formed of a transparent and
relatively rigid thermoplastic material having one or more, in this
case two, cylindrical test chambers 60 and 70 formed extending from
the upper major side 54 toward the lower major side 56. A cartridge
port 53 extends upward from the upper major side 54, and a port
receptacle 55 is sized to receive a test sample dispenser, e.g., a
pipette or a hypodermic needle. Filling channels or capillaries 61
and 71 are machined or molded into the lower major side 56
extending from the port receptacle 55 to the test chambers 60 and
70, respectively. Air vent channels or capillaries 65 and 75 are
also machined or molded into the lower major side 56 extending from
the test chambers 60 and 70, respectively, to air vents 67 and 77,
respectively.
[0079] A sealing sheet 57 is applied against the lower major side
56 in order to close the filling capillaries 61 and 71 and the air
vent capillaries 65 and 75. The sealing sheet is preferably formed
of Mylar.RTM. plastic sheet or polyester film, e.g., ARcare.RTM. 1
mil or 2 mil polyester film available from Adhesives Research,
Inc., of Glen Rock, Pa. The ARcare.RTM. polyester film is coated on
one side with AS-110 acrylic non-migratory inert adhesive that is
applied against the lower major side 56 and does not impede
capillary flow or contaminate the test sample.
[0080] Each air vent 67, 77 may be filled with a vent/fluid seal
plug (not shown) that allows air to escape therethrough while
blocking flow of liquids therethrough as disclosed in the
above-referenced '209 and '286 patents, for example. Suitable
vent/fluid seal plugs made of hydrophobic acrylic copolymer on a
non-woven support or gas permeable films are available from PALL
Life Sciences Division, Ann Arbor, Mich., of Pall Corporation.
Alternatively, a gas permeable film or expanded PTFE fabric layer,
e.g., a GORE-TEX.RTM. laminate filter available from W. L. Gore
& Associates, Inc. may be applied over the upper major surface
54 to extend over and close each air vent 67, 77.
[0081] The closed test chambers 60, 70 of the test cartridge 50
enclosing the test samples protects against exposure to the blood
sample, thereby reducing biohazard risks. The test sample volume is
determined by the volume of the test chambers 60, 70 and is not
dependent upon operator skill to fill the test chamber 60, 70 to a
precise volume. The venting channels and hydrophobic materials
allow venting of air from the test chamber 60, 70 during filling to
enable filling with a precise test volume and eliminates air
bubbles that could alter test results.
[0082] Annular seats 64 and 74 in the upper major surface 54 extend
around the openings of the test chambers 60 and 70, respectively.
Circular chamber caps 62 and 72 formed of a transparent and
relatively rigid thermoplastic material are sized in diameter and
thickness to fit against annular seats 64 and 74 surrounding the
cylindrical test chambers 60 and 70. The peripheral edges of the
chamber caps 62 and 72 are either adhered with adhesive or
thermally bonded by use of ultrasonic welding techniques to the
respective annular seats 64 and 74 to provide a liquid tight seal
of the respective cylindrical test chambers 60 and 70. Each test
chamber 60 and 70 is therefore substantially cylindrical having a
test chamber height between the upper and lower major sides 54 and
56 when the circular chamber caps 62 and 72 are adhered against
annular seats 64 and 74 and a test chamber radius measured from the
chamber pivot element disposed substantially in the center of the
cylindrical test chamber 60 and 70.
[0083] The cylindrical test chambers 60 and 70 further include
chamber pivot elements 68 and 78, respectively, e.g., the depicted
upwardly extending pins or axles that are centrally or axially
disposed within the cylindrical test chambers 60 and 70,
respectively, and extend upward from the lower major side 56. Free
end portions of the chamber pivot elements 68 and 78 are reduced in
diameter to an axle diameter. The lower or inner sides or the
circular chamber caps 62 and 72 are formed with downwardly
extending chamber pivot cups 66 and 76 that are axially aligned
with the centers of the circular chamber caps 62 and 72. The free
ends of the reduced diameter axles of the chamber pivot elements 68
and 78 fit into the downwardly extending chamber pivot cups 66 and
76, respectively, when the circular chamber caps are fitted into
and adhered to the annular seats 64 and 74, respectively.
[0084] It will be understood that the arrangement of the chamber
pivot element 68, 78 with the chamber pivot cup 66, 76 can be
reversed so that the chamber pivot element 68, 78 extends downward
from the respective chamber cap 62, 72, and the chamber pivot cup
66, 76 is formed extending upward into the cylindrical test chamber
60, 70 from the second major side 56. In either case, the reduced
axle diameters of the chamber pivot elements 68 and 78 provide
pivot points of the test chambers 60 and 70 that are intermediate
the upper and lower major sides 54 and 56.
[0085] In the illustrated embodiments, agitators 80 and 100 are
supported by the chamber pivot elements 68 and 78, respectively,
within the respective cylindrical test chambers 60 and 70,
particularly at the reduced axle diameters of the chamber pivot
elements 68 and 78 intermediate the upper and lower major sides 54
and 56. Agitators 80 and 100 comprise agitator pivot elements 88
and 108, respectively, that are engaged by the respective chamber
pivot element 68 and 78. In the depicted embodiment, the agitator
pivot elements 88 and 108 comprise holes or bores sized in bore
diameter to the reduced axle diameters to receive the reduced
diameter portions of the upwardly extending pins or axles of the
respective chamber pivot elements 68 and 78. In this way, the
agitators 80 and 100 are supported at the pivot points of the
respective test chambers 60 and 70 intermediate and spaced from the
upper and lower major sides 54 and 56.
[0086] It will also be understood that the agitator pivot elements
88 and 108 of agitators 80 and 100 could comprise an axle or pin
extending upward to fit into the downwardly extending chamber pivot
cups 66 and 76 and downward to fit into similar pivot cups
extending upwardly from the lower major side 54.
[0087] As called out in FIGS. 4 and 7, the agitators 80 and 100
further comprise an agitator vane formed of agitator vane leaflets
82, 84 and 102, 104, respectively, extending away from agitator
pivot elements 88 and 108, respectively, across the radius of the
respective test chambers 60 and 70 toward the sidewalls thereof.
The agitator vane leaflets 82, 84 and 102, 104 are supported
intermediate the first and second major cartridge sides 54 and 56
by the engagement of the respective agitator pivot elements 88 and
108 with the respective chamber pivot elements 68 and 78.
[0088] The agitator vane leaflets 82 and 84 of agitator 80 extend
away from the agitator pivot element 88 in opposite directions in
substantially a common plane with first and second cut-outs or gaps
86 and 90 separating the agitator vane leaflets 82 and 84, whereby
the agitator 80 has the appearance of a bowtie. Each agitator vane
leaflet 82 and 84 has common upper and lower major surfaces 92 and
94 that are substantially parallel and joined at a vane leaflet
edge or sidewall 96 as shown in FIG. 4.
[0089] Similarly, the agitator vane leaflets 102 and 104 of
agitator 100 extend away from the agitator pivot element 108 in
opposite directions in substantially a common plane with first and
second cut-outs or gaps 106 and 110 separating the agitator vane
leaflets 102 and 104, whereby the agitator 100 has the appearance
of a bowtie. Each agitator vane leaflet 102 and 104 has common
upper and lower major surfaces 112 and 114 that are substantially
parallel and are joined at a vane leaflet edge or sidewall 116 as
shown in FIG. 4.
[0090] As shown in FIG. 6, the agitator 100 is supported within
test chamber 70 at the pivot point comprising the agitator pivot
element 108 and the chamber pivot element 78 to extend
substantially parallel with and between the upper and lower major
sides 54 and 56. The agitator vane leaflets 102 and 104 preferably
each extend from the agitator pivot element 108 substantially
across the test chamber radius. The agitator 80 is supported in
test chamber 60 in the same orientation to the upper and lower
major sides 54 and 56 of the test cartridge housing 52.
[0091] A reagent can be deposited in the blood receptacle 55 and/or
on surfaces of the filling capillaries 61 and 71 and/or the test
chambers 60 and 70, and/or the agitators 80 and 100. A separate
element that is formed of or coated with a reagent can
alternatively or additionally be placed in the test chambers 60 and
70, e.g., in the shape of a washer placed against the base of the
test chambers 60 and 70 below the respective agitators 80 and
100.
[0092] A suitable reagent is selected to react with the blood
sample to facilitate performance of a test on the blood sample
determining test sample coagulation times, e.g., one of TT, PT,
aPTT, and ACT, for example. The reagent is selected from coagulants
or activating agents including calcium, kaolin, celite, ellagic
acid, glass particles, thrombin, thromboplastin or other activating
agents or from heparin neutralizing or deactivating agents
including heparinase or protamine. One test chamber 60 could
contain an activating agent, and the other test chamber 70 could be
devoid of an activating agent or contain a neutralizing agent in
order to perform comparative coagulation time tests.
[0093] Moreover, the agitator vane leaflets 82, 84 and 102, 104 can
be surface etched to increase surface area and to enhance surface
roughness that tends to cause fibrin to develop in blood test
samples as the vane leaflets 82, 84 and 102, 104 are moved through
the blood test samples in the respective test chambers 60 and
70.
[0094] The test chamber 60, 70 may also contain reagents to
counteract any anticoagulants present in the blood sample. During
interventional procedures, heparin may be administered to mitigate
coagulation induced by the procedure, in which case protamine in
the test chamber 60, 70 could counteract heparin and return the
blood sample to a baseline condition. One test chamber 60 could
contain protamine, and the other test chamber 70 could be devoid of
protamine to perform comparative coagulation time tests.
[0095] In accordance with one aspect of the present invention, the
agitators 80 and 100 are rotated or swept about the respective
pivot points in the respective test chambers 60 and 70 to contact
the test sample, particularly a blood test sample, therein. The
agitator vane leaflets 82, 84 and 102, 104 move freely through the
blood test sample until coagulation impedes free movement.
Resistance to agitator movement impedes and signifies coagulation
of the test sample. The reduction of movement of the agitator vane
leaflets 82, 84 and 102, 104 is detected, and coagulation times are
determined for each blood sample. The elapsed time measured from
starting the sweeping movement and the detection of resistance
constitutes a coagulation time.
[0096] In one preferred method, the agitator vane leaflets 82, 84
and 102, 104 are continually swept about the pivot points through
360.degree. revolutions at a predetermined revolution per minute in
an initial mixing phase in order to mix the test sample with a
reagent, if present. A test phase commences upon completion of the
initial mixing phase, wherein the sweeping movement becomes
oscillatory and comprises alternating clockwise and
counter-clockwise sweeps of the agitator vane leaflets 82, 84 and
102, 104 from a neutral position back and forth through
predetermined arcs of less than 360.degree.. The alternating
clockwise and counter-clockwise sweeps of the agitator vane
leaflets 82, 84 and 102, 104 from a neutral angular position are
through predetermined arcs of less than 360.degree., e.g.,
45.degree., 90.degree., or 180.degree. or between 45.degree. and
90.degree. or between 90.degree. and 180.degree.. The coagulation
time is measured either from the commencement of the mixing phase
or the test phase until the resistance to sweeping movement is
detected.
[0097] The resistance caused by the formation of clots or other
increase in viscosity halts or slows agitator vane movement, and
the slowing or absence of sweeping movement is preferably detected
by one or more agitator vane sensor and movement detection
circuitry coupled thereto. The agitator vane leaflet sensor may
comprise one of a proximity sensor, inductive sensor, magnetic
sensor, metal sensor, and optical sensor disposed at a sensor
location with respect to the test chamber.
[0098] The sweeping movement of the agitator vane leaflets 82, 84
and 102, 104 can be effected in many ways. The agitators 80 and 100
are preferably ferromagnetic, formed of stainless steel, for
example, but are not magnetized. Sweeping magnetic fields are
preferably generated that capture the ferromagnetic agitators 80
and 100 and sweep the agitator vane leaflets 82, 84 and 102, 104,
respectively, about a pivot point PP, through the angular positions
depicted in FIGS. 8-12. One of the agitator vane leaflets 82, 84
and 102, 104 is marked for convenience along one outer edge as
shown in FIGS. 8-12 to depict the sweeping movement of each
agitator 80, 100 about the pivot point PP through 360.degree..
[0099] The sweeping magnetic field that effects continuous rotation
of each agitator 80, 100 about a pivot point PP through 360.degree.
revolutions is preferably created by application of four electrical
sweeping signals SS.sub.1, SS.sub.2, SS.sub.3, and SS.sub.4 to four
respective miniaturized electromagnet 42, 44, 46, 48. The four
respective miniaturized electromagnet 42, 44, 46, 48 are supported
within the test instrument 10 arrayed alongside and around each
test chamber 60, 70 when the test cartridge 50 is seated in the
aperture 24. It will be understood that the electromagnets 42, 44,
46, 48 are schematically depicted in FIGS. 8-12, and that the
actual orientation of the physical embodiments may differ from the
depiction. For example, two sets of the physical electromagnets 42,
44, 46, 48 may be disposed in two arrays along one side of the
aperture 24 or distributed for convenience of spacing along both
sides of the aperture 24. The electromagnets 42, 44, 46, 48 may
also comprise electromagnet pairs placed above and below the
aperture 24 receiving the test cartridge 50. Each pair of
electromagnets operating in tandem helps balance the forces applied
to the agitator leaflets or the agitators 80 and 100 and the
sweeping movement about the pivot points within the respective test
chambers 60 and 70.
[0100] The test cartridge housing 52 may also be shaped with
indentations to fit against and dispose the sets of electromagnets
42, 44, 46, 48 in close relation to each test chamber and the vane
leaflets therein so that the primary magnetic flux lines are
aligned in the same plane as the vane leaflets to balance
electromagnetic forces applied to the vane leaflets.
[0101] The sweeping signals SS.sub.1, SS.sub.2, SS.sub.3, and
SS.sub.4 may take any convenient alternating waveform, e.g.,
triangular with a peak as depicted, truncated triangular,
sinusoidal or square. The rise and fall times of each triangular
wave excursion may be the same in slope or may differ. Each
positive excursion of the sweeping signals SS.sub.1, SS.sub.2,
SS.sub.3, and SS.sub.4 causes a magnetic field of a North or South
polarity, and each negative excursion of the sweeping signals
SS.sub.1, SS.sub.2, SS.sub.3, and SS.sub.4 causes a magnetic field
of the opposite South or North polarity. The sweeping signals
SS.sub.1, SS.sub.2, SS.sub.3, and SS.sub.4 are 90.degree. out of
phase with respect to one another. Thus, North and South polarity
magnetic fields are sequentially generated around the circumference
of the test chambers 60 and 70 that pass through the agitator vane
leaflets 82, 84 and 102, 104 and effect rotation of each agitator
80, 90 about its pivot point PP in a clockwise or counterclockwise
(as depicted) direction.
[0102] Generally speaking, each agitator 80, 100 is rotated about
each pivot point PP like the rotor of a brush-less electric motor
as the rising and falling alternating magnetic fields attract and
repel the agitator vane leaflets agitator vane leaflets 82, 84 and
102, 104.
[0103] The sweeping magnetic field that effects back and forth
pivotal movement of the agitators 80 and 100 about each pivot point
PP through an arc less than 360.degree., e.g., about a 180.degree.
arc, is preferably created by application of two of the depicted
alternating sweeping signals SS.sub.1, and SS.sub.3, or SS.sub.2
and SS.sub.4 to the respective miniaturized electromagnets 42 and
46 or 44 and 48. A fixed amplitude, positive polarity, biasing
signal is applied to one of the remaining pair of electromagnets,
and a fixed amplitude, negative polarity, biasing signal is applied
to the other one of the remaining pair of electromagnets. The
relative magnitudes of the sweeping signals as compared to those of
the fixed biasing signals determine the angular displacement of the
vane resulting in an arc of movement between 0.degree. and
180.degree..
[0104] The magnetic field strength is sufficient to selectively
impart sweeping movement of the agitators 80 and 100 through full
360.degree. continuous revolutions and in the back and forth
rotation through about a 180.degree. arc when resistance is low in
the absence of rising viscosity in the test sample, e.g., clotting
or coagulation of a blood sample. But, the magnetic field strength
is not sufficient to overcome resistance to sweeping movement that
develops when such a viscosity increase occurs.
[0105] As shown in FIGS. 8-12 (and in FIG. 15), agitator vane
sensors 40 and 40' are supported by the test instrument 10 along
one side of aperture 24 in relation to each test chamber of a test
cartridge inserted into aperture 24. For example, when the test
cartridge 50 is seated in the aperture 24, each test chamber 60, 70
is disposed alongside a set of agitator vane sensors 40 and 40'. As
shown, one agitator vane sensor 40 is disposed radially aligned
between the pivot point PP and the electromagnet 48, and the second
agitator vane sensor 40' is disposed between adjacent
electromagnets 46 and 48. In practice, either one or both of the
agitator vane sensors 40, 40' can be provided for each test chamber
60, 70.
[0106] The agitator vane sensors 40 and 40' can comprise one of a
proximity sensor, inductive sensor, magnetic sensor, metal sensor,
and optical sensor disposed at an agitator sensor location with
respect to each test chamber 60, 70. The agitator vane sensor 40,
40' for each test chamber 60, 70 provides an agitator sensor output
signal AS OUT upon each sweep of each agitator vane leaflet past
the agitator vane sensor 40, 40'. The AS OUT signals of the
agitator vane sensors 40 and 40' can be processed electrically for
redundancy. Moreover, the AS OUT signals of the agitator vane
sensors 40 and 40' can be processed electrically to obtain
additional information relating to the motion profile of the vane
leaflets moving through the test sample. Changes in acceleration of
the vane leaflets in response to the applied magnetic fields can be
discerned from changes in the phase relationship between the AS OUT
signals generated by the agitator vane sensors 40 and 40'. The
changes in acceleration could potentially indicate clotting
characteristics ("micro" clots, more localized or generalized
clots, etc.) of the test sample prior to the actual clotting event
and indicate further clotting events during the lysis stage after
clotting.
[0107] For convenience, it will be assumed in the following
discussion that only one AS sensor 40 is employed. Preferably, the
agitator vane sensor 40 develops an AS OUT signal exhibiting cyclic
oscillations in amplitude over time as depicted in FIG. 13, for
example, as a function of changes in inductance as the agitator
leaflets sweep past the agitator vane sensor 40. The depicted AS
OUT signal has a period or cycle correlated in time to the applied
sweeping signal and varies in amplitude from a biasing voltage as
the agitator 80, 100 moves freely through the test sample during
the initial phase of a coagulation time test (assuming no failure).
The peak positive and negative amplitude excursions of the AS OUT
signal are relatively large in absolute amplitude with respect to
the baseline 7.9 volts, and the signal cycle length is regular
since the moving agitator vane tracks the changes in the applied
magnetic field.
[0108] An initial AS OUT signal sample window is sampled, digitized
and temporarily stored to be employed as a AS OUT signal template
window TW. As time passes, subsequent AS OUT signal sample windows
SW.sub.1-SW.sub.n are successively defined, sampled, digitized, and
compared to the template window TW employing any simple template
matching algorithm to provide a .DELTA. AS value. The .DELTA. AS
value remains low as long as the current AS OUT signal sample
window SW closely matches the AS OUT signal template window TW,
e.g., in comparing TW with SW.sub.1. The .DELTA. AS value increases
when the current AS OUT signal sample window SW no longer closely
matches the AS OUT signal template window TW, e.g., in comparing TW
with SW.sub.n. Coagulation is declared and the final coagulation
test time is displayed when the .DELTA. AS value increases and
exceeds a .DELTA. AS threshold "T" while the sweeping signals
continue to be applied to the electromagnets 42, 44, 46 and 48 or
pairs thereof. It will be understood that other detection criteria
and algorithms or comparator circuitry could be employed to process
the AS OUT signal and detect slowing or cessation of movement of
the agitator vane.
[0109] A test failure is declared if the .DELTA. AS value increases
and exceeds the .DELTA. AS threshold T during the mixing phase or
during a "too early" initial portion of the test phase. Similarly,
a "maximum test time exceeded" test failure is declared if the
elapsed coagulation test time exceeds a predetermined failure
time.
[0110] It will be understood that tracing of the oscillating AS OUT
signal generated by each agitator vane sensor 40 as illustrated,
for example, in FIG. 13, can also or alternatively be displayed in
the Channel 1 and Channel 2 display lines on LCD screen 26 of test
instrument 10. If such displays are provided, the clinician may
observe the changes that occur in the oscillating AS OUT signal as
fibrin builds up around the agitator vane leaflets, and vane
leaflet motion changes.
[0111] The test cartridge 50 can be employed in the system 10
depicted in FIG. 1 operating in accordance with the steps of the
method depicted in FIG. 14. Certain of the method steps are
practiced by an algorithm and/or circuitry of a preferably
microcomputer-based controller of the test instrument 10.
[0112] Turning to FIG. 14, a test sample, e.g. a blood test sample,
is drawn in step S100 in any conventional manner into a sample
source and is optionally pre-mixed with a reagent. In step S102, a
syringe or a pipette filled with the test sample is directed over
the cartridge port 53 to deposit the test sample into the receiving
chamber 55, and the test sample is conveyed into the test chambers
60 and 70. The test cartridge 50 is inserted in step S104 into the
aperture 24 and may be incubated by the heat block therein to body
temperature for a preset time period as described above. It will be
understood that steps S102 and S104 can be reversed since the test
cartridge 50 could be inserted into the aperture 24 with the
cartridge port 53 exposed so that the test sample can be deposited
into the receiving chamber 55 and passed through the filling
channels into the test chambers 60 and 70.
[0113] As noted above, a mixing phase can be selected by depressing
the Mix switch 20, as determined in step S106, so that the
agitators 80 and 100 are rotated continually for a preset time
period to sweep around and mix the test samples in the respective
test chambers 60 and 70 in step S108. The coagulation test timer is
started at the same time in step S108, since coagulation of the
test sample commences in the mixing phase. Each agitator sensor 40
is also powered in step S110 to derive the AS OUT signals
particularly to ensure that no error condition is present during
the mixing phase. The mixing phase, if selected in step S106, ends
upon time out of a mixing time as determined in step S108.
[0114] When the test phase is commenced in step S110, the agitators
80 and 100 are preferably swept back and forth at a preset rate
through an arc of less than about 180.degree. as described above. A
AS OUT signal template window is derived in step S112 from the AS
OUT signals of each agitator sensor 40 of each channel and
temporarily stored in separate memory registers. Current AS OUT
signal sample windows are successively selected in step S114 as the
AS OUT signals are generated by each agitator sensor 40. The
derived current AS OUT signal sample window of each channel is
compared to the AS OUT signal template window of that channel
employing a template matching algorithm, for example, in step S116
to derive a difference signal .DELTA.AS for the channel. The
difference signal .DELTA.AS of each channel is compared to a
threshold T in step S118.
[0115] For each channel, coagulation is declared and the
coagulation test time is determined in step S120 when the
difference signal .DELTA.AS of the channel exceeds the threshold T
as determined in step S118. The coagulation test time determined in
step S120 is temporarily stored and can be immediately displayed in
the appropriate "Channel 1" or "Channel 2" display line of the LCD
screen 26.
[0116] The coagulation time tests are declared completed in step
S122 when the coagulation times are determined in step S120 for
both blood test samples. Then, it is possible in step S124 to
calculate the average and difference values from the at least two
determined and temporarily stored coagulation test times for
display in the appropriate "Channel 1" or "Channel 2" display line
of the LCD screen 26. The Stop switch 18 is depressed, and the test
cartridge 50 is then removed from the aperture 24 and suitably
disposed of in step S126.
[0117] It will be understood that the algorithm of FIG. 14 may be
modified to provide that the tracings of the oscillating AS OUT
signals generated by each agitator vane sensor 40 are displayed in
the Channel 1 and Channel 2 display lines on LCD screen 26 of test
instrument 10 in addition to the displayed coagulation test times.
If such displays are provided, the clinician may observe the
changes that occur in the oscillating AS OUT signal as fibrin
builds up around the agitator vane leaflets and later due to the
fibrinolytic process. The agitator vane may begin to move again in
response to the applied magnetic fields as fibrinolysis occurs to
weaken the coagulation forces resisting agitator vane movement. The
mechanical integrity of the clot over a longer period of time can
be assessed, particularly to identify coagulopathic characteristics
of the blood test sample before the clinician depresses the Stop
switch 18 in step S126.
[0118] An example of such behavior is depicted in the tracing of AS
OUT signals over a prolonged time period appearing in FIG. 27 that
could be displayed on the LCD screen 26. After an initial 25 second
delay, the AS OUT signal varies between the baseline 7.9 volts and
about 4.6 volts until about 75 seconds have elapsed. Then, the AS
OUT signal variation diminishes to between the baseline 7.9 volts
and about 6.0 volts due to a reduction in vane movement as movement
is resisted by coagulation of the test sample. Coagulation would be
declared at about 80 seconds employing the techniques described
above. Then, the AS OUT signal varies between the baseline 7.9
volts and about 5.5 seconds at about 220 seconds as the agitator
vane begins to move more freely in response to the applied magnetic
fields and as fibrinolysis occurs to weaken the coagulation forces
resisting agitator vane movement. At about 350 seconds, coagulation
and fibrinolysis re-balance is restored, as evidenced by the
resumption of variation of the AS OUT signal between the baseline
7.9 volts and about 4.6 volts. Thus, the clinician is afforded a
more complete understanding of the coagulopathic characteristics of
the blood test sample. This long term behavior of the blood test
sample depicted, for example in the tracing of FIG. 27 is not
possible to observe with prior art coagulation timers that depend
upon devices that passively fall under the force of gravity through
the test sample.
[0119] In the depicted preferred embodiments described above, two
test chambers 60 and 70 are included in each test cartridge 50, and
the instrument 10 is configured and programmed to simultaneously
perform redundant coagulation time tests to provide increased
accuracy or distinct coagulation time tests of test samples exposed
to differing reagents to derive distinct comparative coagulation
times. Other embodiments of the test cartridge may include
additional test chambers to increase redundancy or to perform
differing tests on the same test sample simultaneously or
sequentially, and the test instrument may be modified to perform
the additional tests.
[0120] For example, six test chambers can be included in each test
cartridge in an array that is adapted to be inserted into the
aperture of a test instrument having six sets of internally
disposed agitator rotating electromagnet sets and agitator sensors
operable substantially as described herein. In such a six-chamber
test cartridge, three redundant tests can be conducted employing
differing reagents. For example, two test chambers can be devoid of
any reagent, two test chambers can include a small quantity of
heparin, and two further test chambers can include a larger
quantity of heparin. The three duplicate ACT test results provide
heparin dose response (HDR) data points that can be plotted to
determine a patient's individual heparin response. The patient's
heparin needs during bypass can be extrapolated based on this
response curve.
[0121] The above described agitators 80 and 100 can be
characterized as "flat" agitators because the agitator sidewalls 96
and 116 are relatively small in surface area compared to the upper
and lower major surfaces 92, 94 and 112, 114, respectively.
Moreover, the agitator thickness, i.e., the width or height of the
sidewalls 96 and 116, is relatively small in relation to the depth
or height (hereafter test chamber height) of the test chambers 60
and 70 as shown in FIG. 6. The sidewalls of the flat agitator
leaflets 82, 84 and 102, 104 present agitator sweeping face surface
areas applied against the test sample that are about less than 40%
of the test chamber cross-section areas of the respective test
chambers 60 and 70. While excellent coagulation time tests can be
performed on blood and biologic test samples, there may be concern
that coagulation times are prolonged because the agitator sweeping
faces sweeping against less than 50% of the volume of the test
samples may tend to cut through developing fibrin strands and
clots.
[0122] In accordance with a further aspect of the present
invention, alternative agitators *80, schematically depicted in
FIGS. 15 and 16, have been developed and tested that are shaped to
have agitator vane leaflets having agitator sweeping faces that
sweep through a higher fraction of the test chamber height during
rotation of the agitator vane leaflets. The schematically depicted
agitator *80 comprises first and second agitator vane leaflets *82
and *84 extending in opposed directions away from the agitator
pivot elements *88 and *90. It will be understood that the agitator
pivot elements *88 and *90 fit within cup-shaped test chamber pivot
elements of a test chamber 160 of a modified test cartridge 150,
whereby the agitator and test chamber pivot elements together
define the test chamber pivot point PP and a pivot point axis PP-PP
as shown in FIG. 16. It will also be understood that the
alternative pivot point PP and pivot axis provided by the axle and
hole structure depicted in the test cartridge 50 in FIGS. 3, 5 and
6 can be substituted for the pivot elements *88 and *90 fitting
within the cup-shaped test chamber pivot elements.
[0123] As shown in FIG. 16, the agitator vane and the vane leaflets
*82 and *84 have an agitator vane height AVH, and the agitator vane
leaflets *82 and *84 have a leaflet thickness, and a vane leaflet
length VLL measured from the pivot point axis at PP-PP. Each
agitator vane leaflet *82 and *84 extends from the pivot point PP
substantially across the test chamber radius TCR. The agitator vane
length AVL is equal to twice the vane leaflet length VLL, and the
test chamber diameter TCD is equal to twice the test chamber radius
TCR. The agitator vane height AVH is somewhat smaller than the test
chamber height TCH, and the leaflet thickness is less than the
agitator vane height AVH and the vane leaflet length VLL. The vane
leaflet sweeping faces have a surface area defined by AVH.times.AVL
that can be up to about 90% of the test chamber cross-section area
defined by TCH.times.TCD. Stated another way, the agitator vane of
agitator *80 has an agitator vane height AVH extending
substantially through the test chamber height TCH and an agitator
vane length AVL extending substantially through the test chamber
diameter TCD.
[0124] The enlarged leaflet sweeping faces *92 and *94 present high
fluid interface profiles to the fluid test sample as the fluid test
sample washes against the leaflet sweeping faces *92 and *94 during
the sweeping movement of the agitator vane leaflets *82 and *84
about the pivot point PP and through the test sample within test
chamber 160.
[0125] In certain preferred embodiments, the leaflet sweeping faces
*92 and *94 preferably are shaped as paddles wherein the respective
leaflet sweeping faces *92 and *94 are substantially parallel to
one another and are substantially parallel to the pivot axis as
shown in FIGS. 15 and 16. In other embodiments, the leaflet
sweeping faces *92 and *94 of the agitator vane leaflets *82 and
*84 are not necessarily parallel to one another, but are symmetric.
The agitator embodiments of FIGS. 17-26 are denoted *80, wherein *
is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10; each such agitator embodiment can
be substituted for the flat agitators 80 and 100 of the test
cartridge 50 or the modified test cartridge 150 depicted in FIG. 17
(with suitable agitator and test chamber pivot elements).
[0126] A first alternative agitator 180 is depicted in FIG. 17 in
relation to the modified test cartridge 150 having a single test
chamber 160 that is formed in a cartridge housing 152 and enclosed
by a circular chamber cap 162. The cartridge housing 152 has a
substantially prismatic shape with upper and lower major sides 154
and 156 and a minor cartridge edge or sidewall 158 extending
between the upper and lower major sides 154 and 156 and maintaining
the upper and lower major sides 154 and 156 in substantially
parallel relation. Finger grip indentations can be formed in the
minor cartridge sidewall 158 to facilitate holding the test
cartridge 150 upright, filling test chambers with the test sample,
inserting and removing the cartridge housing 152 into and from the
instrument cartridge receptacle 24. A port receptacle 155 into the
upper major side 154 is sized to receive a test sample dispenser,
e.g., a pipette or a hypodermic needle. A filling channel is
machined or molded into the lower major side 156 extending from the
port receptacle 155 to the test chamber 160. An air vent channel
165 is also machined or molded into the lower major side 56
extending from the test chamber 160 to an air vent 167 that may be
sealed as described above with respect to air vent 167. A sealing
sheet can be applied against the lower major side 156 in order to
close the filling capillary and the air vent capillary 165. An
annular seat 164 in the upper major surface 154 extends around the
opening of the test chamber 160. The circular chamber cap 162 is
formed of a transparent and relatively rigid thermoplastic material
that is are sized in diameter and thickness to fit against and be
sealed into annular seat 164 as described above with respect to
chamber caps 62 and 72. One cup-shaped test cartridge pivot element
168 is depicted in FIG. 17. The test cartridge 150 may have one or
more additional test chamber as described above with respect to the
test cartridge 50.
[0127] In each of the following described embodiments of the
agitator *80, the change in agitator vane movement is detected from
the processing of the AS OUT signals developed by the agitator
sensor 40 as described above with respect to FIGS. 13 and 14.
[0128] The alternative agitator 180 depicted in FIG. 17 comprises
agitator vane leaflets 182 and 184 extending away from vane pivot
elements 188 and 190 that have respective leaflet sweeping faces
192 and 194. The vane leaflets 182 are shaped so that the
respective leaflet sweeping faces 192 and 194 are curved or concave
like a scoop along the vane leaflet length VLL from the pivot point
PP defined by vane pivot elements 188 and 190 as shown in FIG. 15.
In blood test samples, fibrin strands forming during coagulation
are scooped up, slowing rotation or reducing the agitator vane
range of motion, and possibly eventually halting the rotation of
the agitator 180 about the pivot point PP.
[0129] The alternative "window" agitator 380 depicted in FIG. 18
has a rectangular frame 396 defining a window supported by the
agitator pivot elements 388 and 390 to provide the opposed vane
leaflets 382 and 384. The opposed vane leaflets 382 and 384 have
vane leaflet sweeping faces 392 and 394 that sweep through the test
sample in test chamber 160 when the agitator vane leaflets 382 and
384 are rotated about the pivot point PP in the counterclockwise
direction as shown in FIG. 15. In blood test samples, fibrin
strands forming during coagulation catch on the rectangular frame
396 tending to fill the window and slowing rotation or reducing the
agitator vane range of motion, and possibly eventually halting the
rotation of the window agitator 380 about the pivot point PP.
[0130] The alternative "mesh" agitator 480 depicted in FIG. 19 has
porous mesh layers 492 and 494 supported by or adhered to the sides
of the hollow rectangular frame forming the opposed vane leaflets
482 and 484 supported by the agitator pivot elements 488 and 490.
The porous mesh layers 492 and 494 can comprise wire or fabric mesh
layers or a single porous mesh layer fitted within the rectangular
frame. The porous mesh layers 492 and 494 constitute the vane
leaflet sweeping faces that sweep through the test sample in test
chamber 160 when the agitator vane leaflets 482 and 484 are rotated
about the pivot point PP in the counterclockwise direction as shown
in FIG. 15. In blood test samples, fibrin strands forming during
coagulation catch on the mesh layers 492 and 494, slowing rotation
or reducing the agitator vane range of motion, and possibly
eventually halting the rotation of the mesh agitator 480 about the
pivot point PP.
[0131] The opposed vane leaflets 582 and 584 of the "comb" agitator
580 supported by the agitator pivot elements 588 and 590 are
slotted with slots 596 as depicted in FIG. 20 to form a plurality
of comb tines that sweep through the test sample. The opposed vane
leaflets 582 and 584 have vane leaflet sweeping faces 592 and 594,
respectively, that sweep through the test sample in test chamber
160 when the agitator vane leaflets 582 and 584 are rotated about
the pivot point PP in the counterclockwise direction as shown in
FIG. 15. In blood test samples, fibrin strands forming during
coagulation are raked out of the test sample by the tines and
accumulate in the slots 596, slowing rotation or reducing the
agitator vane range of motion, and possibly eventually halting the
rotation of the comb agitator 580 about the pivot point PP.
[0132] Sieve" holes 696 are formed as depicted in FIG. 21 extending
through the opposed vane leaflets 682 and 684 of the sieve agitator
680 supported by the agitator pivot elements 688 and 690 that sweep
through the test sample. The opposed vane leaflets 682 and 684 have
vane leaflet sweeping faces 692 and 694, respectively, that sweep
through the test sample in test chamber 160 when the agitator vane
leaflets 682 and 684 are rotated about the pivot point PP in the
counterclockwise direction as shown in FIG. 15. In blood test
samples, fibrin strands forming during coagulation catch and
accumulate in the sieve holes 696, slowing rotation or reducing the
agitator vane range of motion, and possibly eventually halting the
rotation of the sieve agitator 680 about the pivot point PP.
[0133] Notches 796 are formed as depicted in FIG. 22 along the
upper and lower edges of the opposed vane leaflets 782 and 784 of
the "notched bar" agitator 780 supported by the agitator pivot
elements 788 and 790 that sweep through the test sample. The
opposed vane leaflets 782 and 784 have vane leaflet sweeping faces
792 and 794, respectively, that sweep through the test sample in
test chamber 160 when the agitator vane leaflets 782 and 784 are
rotated about the pivot point PP in the counterclockwise direction
as shown in FIG. 15. In blood test samples, fibrin strands forming
during coagulation catch and accumulate in the notches 796, slowing
rotation or reducing the agitator vane range of motion, and
possibly eventually halting the rotation of the notched bar
agitator 780 about the pivot point PP.
[0134] Notches 896 and 898 are formed as depicted in FIG. 23 along
the vane leaflet ends as well as the upper and lower edges of the
opposed vane leaflets 882 and 884 of the "notched end" agitator 880
supported by the agitator pivot elements 888 and 890 that sweep
through the test sample. The opposed vane leaflets 882 and 884 have
vane leaflet sweeping faces 892 and 894, respectively, that sweep
through the test sample in test chamber 160 when the agitator vane
leaflets 882 and 884 are rotated about the pivot point PP in the
counterclockwise direction as shown in FIG. 15. In blood test
samples, fibrin strands forming during coagulation catch and
accumulate in the notches 896 and 898, slowing rotation or reducing
the agitator vane range of motion, and possibly eventually halting
the rotation of the notched agitator 880 about the pivot point
PP.
[0135] A "tube" agitator 980 is depicted in FIG. 24 where the
agitator vane comprises an elongated tube having a tube diameter
corresponding to agitator height AVH of FIG. 16 and forming the
agitator vane leaflets 982 and 984 extending away from the agitator
pivot element 988. The agitator pivot element 988 in this example
comprises an axle bore extending transverse to the tube axis
adapted to receive an axle, centrally or axially disposed within
the cylindrical test chamber 160, e.g., the axle 78 of the test
chamber 70 as depicted in FIG. 6 and described above. The opposed
vane leaflets 982 and 984 have vane leaflet sweeping faces 992 and
994, respectively, that present symmetric convex curved leaflet
sweeping faces against the test sample disposed in the test chamber
during rotation of the agitator vane leaflets 982 and 984. The vane
leaflet sweeping faces 992 and 994 sweep through the test sample in
test chamber 160 when the agitator vane leaflets 982 and 984 are
rotated about the pivot point PP in the counterclockwise direction
as shown in FIG. 15. In blood test samples, fibrin strands forming
during coagulation catch and accumulate around the convex vane
leaflet sweeping faces 992 and 994, slowing rotation or reducing
the agitator vane range of motion, and possibly eventually halting
the rotation of the tube agitator 880 about the pivot point PP.
[0136] A "U-shape" agitator 1080 is depicted in FIG. 25 where the
agitator vane comprises a U-shaped channel having a channel side
height corresponding to agitator height AVH of FIG. 16 and forming
the agitator vane leaflets 1082 and 1084 extending away from the
agitator pivot element 1088. The elongated U-shaped channel extends
between opposed channel ends and an agitator pivot element 1088 is
formed between the opposed channel ends. The agitator pivot element
1088 in this example comprises an axle bore extending transverse to
the tube axis adapted to receive an axle, centrally or axially
disposed within the cylindrical test chamber 160, e.g., the axle 78
of the test chamber 70 as depicted in FIG. 6 and described above.
The opposed vane leaflets 1082 and 1084 have vane leaflet sweeping
faces 1092 and 1094, respectively, that sweep through the test
sample in test chamber 160 when the agitator vane leaflets 1082 and
1084 are rotated about the pivot point PP in the counterclockwise
direction as shown in FIG. 15. In blood test samples, fibrin
strands forming during coagulation catch and accumulate on the vane
leaflet sweeping faces 1092 and 1094, slowing rotation or reducing
the agitator vane range of motion, and possibly eventually halting
the rotation of the U-shape agitator 1080 about the pivot point
PP.
[0137] A "propeller" agitator 280 is depicted in FIG. 26 wherein
the agitator vane comprises a propeller having opposed propeller
blades each having a propeller blade height corresponding to
agitator height AVH of FIG. 16. The agitator pivot element 288 in
this example comprises an axle bore extending transverse to the
tube axis adapted to receive an axle, centrally or axially disposed
within the cylindrical test chamber 160, e.g., the axle 78 of the
test chamber 70 as depicted in FIG. 6 and described above. The
agitator vane leaflets 282 and 284 extending away from the agitator
pivot element 288 are shaped like symmetric propeller blades having
complementary pitches with respect to the pivot axis. The propeller
blade shaped vane leaflets 282 and 284 therefore have vane leaflet
sweeping faces 292 and 294, respectively, that sweep through the
test sample in test chamber 160 when the agitator vane leaflets 282
and 284 are rotated about the pivot point PP in the
counterclockwise direction as shown in FIG. 15. In blood test
samples, fibrin strands forming during coagulation catch and
accumulate on the vane leaflet sweeping faces 1092 and 1094,
slowing rotation or reducing the agitator vane range of motion, and
possibly eventually halting the rotation of the propeller agitator
280 about the pivot point PP.
[0138] Thus, the leaflet sweeping faces can be solid and
impermeable to the test sample or can be permeable to the test
sample. The agitator vane leaflets can have leaflet sweeping faces
and edges that are relatively smooth or are notched or roughened.
The leaflet sweeping faces may also be coated or roughened to
encourage coagulation.
[0139] As noted above, the agitator vane sensor continuously
monitors agitator vane movement thereby capturing any change in
movement as soon as the change occurs. In this way, there is no lag
time between a change in agitator vane movement and detection of
the change.
[0140] All patents and publications referenced herein are hereby
incorporated by reference in their entireties.
[0141] It will be understood that certain of the above-described
structures, functions and operations of the above-described
preferred embodiments are not necessary to practice the present
invention and are included in the description simply for
completeness of an exemplary embodiment or embodiments.
[0142] In addition, it will be understood that specifically
described structures, functions and operations set forth in the
above-referenced patents can be practiced in conjunction with the
present invention, but they are not essential to its practice.
[0143] It is to be understood, that within the scope of the
appended claims, the invention may be practiced otherwise than as
specifically described without actually departing from the spirit
and scope of the present invention. The disclosed embodiments are
presented for purposes of illustration and not limitation, and the
present invention is limited only by the claims that follow.
* * * * *