Capillary flow device and method for measuring activated partial thromboplastin time

• US 5,039,617 A
• Filed: 04/20/1989
• Issued: 08/13/1991
• Est. Priority Date: 04/20/1989
• Status: Expired due to Term

• Current Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Original Assignee: Biotrack, Inc. (Novartis Ag)
• Inventors: Hill, James L., Gorin, Michael M., McDonald, Katheryne M., Hsu, Po C., Winfrey, Laura J.
• Primary Examiner(s): Lacey, David L.
• Assistant Examiner(s): Snay, Jeffrey R.
• Application Number:
  US07/341,045
• Time in Patent Office: 845 Days
• Field of Search: 436/69, 436/180, 435/13, 435/810, 422/57, 422/58, 422/68, 422/73, 422/81, 422/82
• US Class Current: 436/69
• CPC Class Codes: G01N 33/4905 Determining clotting time of blood by chemical methods G01N33/86, C12Q1/54
• CPC Class Codes: G01N 33/86 involving blood coagulating time or factors, or their receptors
• CPC Class Codes: Y10S 435/81 Packaged device or kit

Abstract

Methods and devices are provided for carrying out activated partial thromboplastin time (APTT) analysis on a whole blood sample to which no anticoagulant has been added by applying the sample to a capillary track contained in a housing, wherein the capillary track comprises sample entry and venting ports, at least one capillary track connecting the entry and venting ports, and reagents capable of initiating an APTT analysis, wherein clotting time is measured by the cessation of blood flow in the capillary track.

Claims

1. In a method for determining activated partial thromboplastin time in a fluid medium employing a device comprising, in a housing, a continuous capillary track comprising an inlet port, at least one capillary unit acting as the motive force for moving the fluid medium in the device, and a venting port, wherein said capillary track further contains a reagent capable of initiating blood clotting, and in which a blood sample is introduced into said entry port, said sample is allowed to transit said capillary track by capillary action, and said activated partial thromboplastin time is determined from the time required for blood coagulation and flow stoppage in said capillary track, the improvement that comprises:

utilizing as said reagent a mixture of (1) an activating agent for activated partial thromboplastin time measurements wherein said activating agent is a sulfatide or sulfoglycosylsphingolipid and (2) a phospholipid component, said activating agent and said phospholipid component being provided in amounts sufficient to initiate clotting, and to provide a time of flow stoppage that is essentially independent of blood hematocrit.

2. The method of claim 1, wherein said phospholipid component comprises a compound selected from the group consisting of soybean phospholipid type II-S, rabbit brain cephalin, rabbit lung cephalin, human brain cephalin, and a mammalian platelet extract.

3. The method of claim 1, further comprising providing said capillary track to allow continuous flow of whole blood in said capillary track in the absence of said reagent for at least 200 seconds.

4. The method of claim 3, wherein said continuous flow is achieved using no more than 40 μ

l of sample.

5. The method of claim 1, wherein said capillary track comprises a reagent unit and a capillary flow unit exiting said reagent unit.

6. The method of claim 5, wherein at least a portion of said capillary flow unit has a cross sectional area that increases with increasing distance from said reagent unit.

7. The method of claim 5, wherein said capillary flow unit has an initial cross sectional area of no more than 0.05 mm².

8. The method of claim 1, wherein said blood sample is whole blood.

9. The method of claim 8, wherein no chemical treatment of said blood takes place between obtaining said blood from a subject and applying said blood to said inlet port.

10. The method of claim 8, wherein movement of red blood cells past a point in said capillary track is detected by measurement of light scatter.

11. The method of claim 8, wherein said blood sample is capillary blood.

12. The method of claim 11, wherein said blood sample has a volume of no more than about 40 μ

l.

13. In a device for determining activated partial thromboplastin time comprising, in a housing, a continuous capillary track comprising an inlet port, at least one capillary unit acting as the motive force for moving the fluid medium in the device, and a venting port, wherein said capillary track further contains a reagent capable of initiating blood clotting, and in which a blood sample is introduced into said entry port, said sample is allowed to transit said capillary track by capillary action, and said activated partial thromboplastin time is determined from the time required for blood coagulation and flow stoppage in said capillary track, the improvement that comprises:

provided as said reagent a mixture of (1) an activating agent for activated prothrombin time measurements wherein said activating agent is a sulfatide or sulfoglycosylsphingolipid and (2) a phospholipid component, said activating agent and said phospholipid component being provided in amounts sufficient to initiate blood clotting, and to provide a time of flow stoppage that is essentially independent of blood hematocrit.

14. The device of claim 13, wherein said phospholipid component comprises a compound selected from the group consisting of soybean phospholipid type II-S, rabbit brain cephalin, rabbit lung cephalin, human brain cephalin, and mammalian platelet extract.

15. The device of claim 13, wherein said capillary track allows continuous flow of whole blood in said capillary track in the absence of said reagent for at least 200 seconds.

16. The device of claim 15, wherein said continuous flow is achieved using no more than 40 μ

l of sample.

17. The device of claim 13, wherein said capillary track comprises a reagent unit and a capillary flow unit exiting said reagent unit.

18. The device of claim 17, wherein at least a portion of said capillary flow unit has a cross sectional area that increases with increasing distance from said reagent unit.

19. The device of claim 17, wherein said capillary flow unit has a initial cross sectional area of no more than 0.05 mm².

20. The device of claim 13, wherein said device comprises first and second plastic plates welded together and said capillary track is formed between said plates and comprises a depression in a welded surface of said first plate, the second plate being flat at the location of said capillary track.

21. The device of claim 20, wherein said depression is substantially semicircular in the region of said capillary flow unit.

22. A capillary flow device capable of being used in an analysis for activated partial thromboplastin time, comprising:

a housing containing;

a continuous capillary pathway comprising an entry port, a first capillary unit, a reagent chamber, a second capillary unit, and a venting port; and

a reagent in said reagent chamber, wherein;

(1) said first capillary unit comprises a capillary channel from 5 to 15 mm in length and having a width in the range of from 0.75 to 2 mm, and a height in the range of from 0.05 to 0.125 mm;

(2) said reagent chamber comprises a capillary chamber having a length in the range of from 5 to 15 mm, a width in the range of from 3 to 8 mm, and a height in the range of from 0.05 to 0.125 mm;

(3) said second capillary unit comprises a capillary channel having an initial radius in the range of from 0.05 to 0.175 mm and a final radius that provides capillary flow for a blood sample, said channel having one flat side, the remaining sides of said channel being formed by flat surfaces or curved surfaces joined without forming angles, all curved surfaces being convex relative to said flat side, wherein at least a portion of said second capillary increases in effective diameter with increasing distance from said reagent chamber;

(4) said reagent comprises an activator for activated partial thromboplastin time wherein said activator is a sulfatide or sulfoglycosylsphingolipid and a phospholipid component said activator and said phospholipid component being provided in amounts sufficient to initiate blood clotting and to provide a time of flow stoppage that is essentially independent of blood hematocrit clotting; and

(5) said capillary track having a total volume in the range of from 20 to 100 μ

l.

23. The device of claim 22, wherein said second capillary unit has a volume of no more than 40 μ

l.

24. The device of claim 22, wherein said second capillary unit has a volume of no more than 25 μ

l.

25. The device of claim 22, wherein said housing comprises at least two plastic, pieces joined together and said second capillary unit is formed between a first plastic piece that is flat in the region of said second capillary unit and a second plastic piece that has a semicircular depression in the region of said second capillary unit.

26. The device of claim 25, wherein said semicircular depression has a constant radius for a first portion of said capillary unit.

27. The device of claim 26, wherein said semicircular depression has a radius that increases in a second portion following said initial portion.

28. The device of claim 27, wherein said second portion has a radius that increases with a decreasing rate of increase.