Analytical test device and method for use in medical diagnoses

• US 6,171,870 B1
• Filed: 08/06/1998
• Issued: 01/09/2001
• Est. Priority Date: 08/06/1998
• Status: Expired due to Fees

• Current Assignee: Nexus Dx, Inc. (Sphingotec GMBH)
• Original Assignee: Spectral Diagnostics, Inc.
• Inventors: Freitag, Helmut E.
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): PHAM, MINH QUAN K
• Application Number:
  US09/130,164
• Time in Patent Office: 887 Days
• Field of Search: 435/5, 435/7.1, 435/7.2, 435/7.24, 435/7.29, 435/7.92, 435/287.2, 435/288.2, 435/6, 435/91.2, 435/4, 435/7.5, 435/7.9, 436/514, 436/516, 436/518, 436/164, 436/165, 436/69, 422/56, 422/57, 422/58, 422/61, 422/72, 422/82.06, 250/341
• US Class Current: 436/518
• CPC Class Codes: G01N 2333/4712 Muscle proteins, e.g. myosin, actin, protein
• CPC Class Codes: G01N 2333/805 Haemoglobins; Myoglobins
• CPC Class Codes: G01N 2333/9123 with a nitrogenous group as acceptor (2.7.3), e.g. histidine kinases
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/54389 with bidirectional or multidirectional lateral flow, e.g. wherein the sample flows from a single, common sample application point into multiple strips, lanes or zones
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387

Abstract

An analytical test device is described for the immunochromatographic determination of the presence of one or more analytes in fluid samples. The device is configured such that the sample is allowed to enter the detection zone simultaneously from many different directions, eliminating stagnation of the flow of the sample. By selection of the porous substrate, the device also allows for the separation of red blood cells from plasma, providing a rapid test for one or more analytes in a sample of whole blood. The device of the present invention may measure more than one analyte simultaneously from a single sample, either by having multiple immunochromatographic pathways fed by a single sample, or multiple analytes detected in the same pathway by way of multiple capture antibodies.

Claims

1. An analytical test device for determining the presence of at least one analyte in a fluid sample, said device comprising:

a dry porous carrier;

at least one detection zone covering at least a segment of an area of said carrier;

a sample circulation channel in operative communication with said detection zone, the sample being permitted to enter from said sample circulation channel into said detection zone from a plurality of different directions;

at least one capture zone channel having an entrance end which is in operative communication with the detection zone to permit sample to flow from the detection zone into the capture zone channel, the distances between all points where the sample is permitted to enter the detection zone and said entrance end being essentially the same.

2. The device of claim 1 containing reagents required for reactions determinative of the analyte.

3. The device of claim 1 wherein said area is semicircular.

5. The device of claim 1 or 4 wherein the analyte is selected from troponin I, troponin T, myoglobin, CK-MB and mixtures thereof.

6. The device of claim 1 or 4 wherein the analyte is selected from hCG, LH and mixtures thereof.

7. The device of claim 1 or 4 wherein the fluid sample is selected from whole blood, plasma, serum and urine.

8. The device of claim 1 or 4 wherein the dry porous carrier is nitrocellulose.

9. The device of claim 1 or 4 comprising two or more detection zones arranged on said area of the carrier, there being an equal number of capture zone channels, each in operative communication with a corresponding detection zone.

11. The device of claim 1 or 4 wherein said area is polygonal or forms part of a polygon.

12. The device of claim 11 wherein said area is rectangular or forms part of a rectangle.

4. An analytical test device for determining the presence of at least one analyte in a fluid sample, said device comprising:

a dry porous carrier through which a sample can flow by capillary action;

a sample delivery means through which the sample can be applied to the device and flow thereinto;

a sample circulation channel which is closed at its terminal ends and which circumscribes an area of the carrier;

at least one detection zone covering at least a segment of said area of the carrier, wherein at least a segment of the sample circulation channel conforms with a border of the detection zone and is in operative communication with said detection zone, across which border the sample is permitted to enter into the detection zone simultaneously from a plurality of different directions and to form a stream flowing away from said border, the detection zone containing at least one mobile, labelled specific binding reagent for the at least one analyte, which said at least one mobile, labelled specific binding reagent is capable of reacting with said at least one analyte to form at least one labelled complex which is capable of moving along with said stream; and

at least one capture zone channel having an entrance end and a closed terminal end, the entrance end being in operative communication with the detection zone to permit said stream to flow from the detection zone into the capture zone channel, the distances between all points of said border and said entrance end being essentially the same, the capture zone channel containing at least one immobilized specific binding reagent, which at least one immobilized specific binding reagent is capable of reacting with and concentrating said at least one labelled complex to form at least one detectable reaction product.

10. The device of claim 4 wherein the sample circulation channel is arcuate and said area is semicircular.

13. The device of claim 4 wherein said sample circulation channel is polygonal or forms part of a polygon.

14. The device of claim 13 wherein said sample circulation channel is rectangular or forms part of a rectangle.

15. The analytical test device of claim 4 suitable for determining the presence of at least one analyte contained in a liquid sample while permitting rapid and efficient flow of the sample through at least one defined pathway in which reactions determinative of the analyte take place, said device comprising:

a top support layer and a bottom support layer, the top support layer having an upper surface and a lower surface, the upper surface being formed with a through hole for the addition of the sample, the through hole being in registry with a sample delivery channel formed in the lower surface of the top layer and in operative communication with a sample circulation channel closed at its terminal ends and formed with inner and outer walls to define a pathway for the sample in the lower surface of the top layer, the inner walls of the sample circulation channel defining an indent in the lower surface of the top layer;

the bottom support layer being attached to the top layer;

a dry porous carrier layer through which the sample can flow by capillary action sandwiched between the top support layer and the bottom support layer, the porous carrier layer being configured to contain at least one detection zone having a border in operative communication with a segment of the sample circulation channel and a second opposite end in operative communication with a capture zone channel having an entrance end and a closed terminal end thereby to provide a conduit through which the liquid sample may flow by capillary action from the sample circulation channel to the terminal end of the capture zone channel;

the sample delivery channel, sample circulation channel, detection zone and capture zone channel forming a defined pathway through which the liquid sample flows from the through hole to the terminal end of the capture channel;

there being at least one mobile, labelled reagent which will specifically react with the analyte in the detection zone to form a labelled complex which will move by capillary action into the capture zone channel; and

there being immobilized reagent in the capture zone channel which will react with and concentrate the labelled complex to form a detectable reaction product.

16. The device of claim 15 in which the sample circulation channel is arcuate.

17. The device of claim 15 in which the liquid sample is selected from whole blood, plasma, serum and urine.

18. The device of claim 15 wherein the analyte is selected from troponin I, troponin T, myoglobin, CK-MB and mixtures thereof.

19. The device of claim 15 wherein the analyte is selected from hCG, LH and mixtures thereof.

20. The device of claim 15 wherein the upper layer is transparent and the detectable reaction product is visible.

21. The device of claim 15 wherein the upper layer is opaque and has a view window through which the detectable reaction product is visible.

22. The device of claim 15 wherein said at least one analyte is a cardiac analyte and said liquid sample is whole blood containing red blood cells and plasma, the dry porous carrier causing said red blood cells to separate chromatographically from the plasma to provide a plasma front moving in the capture zone channel and a red blood cell front upstream thereof, a detectable portion of the labelled complex being between the red blood cell front and the plasma front so that the labelled complex is substantially free of red blood cells when said labeled complex is captured in the capture channel by reaction with the immobilized reagent.

23. The device of claim 22 wherein the mobile reagent in the detection zone is a labelled antibody which will react with one epitope on the analyte and the immobilized reagent is an antibody which will react with another epitope on the analyte.

24. The device of claim 22 wherein, in the detection zone, there is a mixture of a mobile labelled antibody which will react with one epitope on the analyte and a mobile biotin labelled antibody which will react with another epitope on the analyte, and immobilized reagent is an avidin.

25. The device of claim 22 containing one defined pathway for determining the presence of troponin I or troponin T.

26. The device of claim 22 containing two defined pathways, one for determining the presence of myoglobin, the other for determining the presence of CK-MB.

27. The device of claim 22 containing three defined pathways, the first pathway for determining the presence of troponin I or troponin T, the second pathway for determining the presence of myoglobin and the third pathway for determining the presence of CK-MB.

28. The device of claim 27 containing one defined pathway for determining the presence of troponin I.

29. The device of claim 22 in which there is only one defined pathway and it contains the reagents for determining the presence of myoglobin and CK-MB.

30. The device of claim 22 containing one defined pathway for determining the presence of troponin I or troponin T, the presence of myoglobin and the presence of CK-MB.

31. The device of claim 15 wherein, in the detection zone, there is a mixture of a mobile labelled antibody which will react with one epitope on the analyte and a mobile biotin labelled antibody which will react with another epitope on the analyte, and the immobilized reagent is selected from the group consisting of streptavidin and avidin.

32. The device of claim 15 wherein the label is a particulate direct label.

33. The device of claim 32 wherein the label is a gold label.

34. The device of claim 15 in which the dry porous carrier is nitrocellulose.

35. The device of claim 15 wherein said area is polygonal or forms part of a polygon.

36. The device of claim 35 wherein said area is rectangular or forms part of a rectangle.

37. A method for determining the presence of at least one analyte in a fluid sample comprising the sequential steps of:

i) providing an analytical test device suitable for determining the presence of at least one analyte contained in a liquid sample while permitting rapid and efficient flow of the sample through at least one defined pathway in which reactions determinative of the analyte take place, said device comprising;

a) a dry porous carrier through which sample can flow by capillary action;

b) at least one detection zone covering at least a segment of an area of said carrier, and a sample circulation channel in operative communication with said detection zone, such that said sample is permitted to enter from said sample circulation channel into said detection zone from a plurality of different directions, and said detection zone containing at least one mobile, labelled binding reagent for said at least one analyte, which mobile, labelled binding reagent is capable of reacting with said analyte to form a labelled complex;

c) at least one capture zone channel having an entrance end which is in operative communication with the detection zone to permit sample to flow from the detection zone into the capture zone channel, the distances between all points where said sample is permitted to enter said detection zone and said entrance end being essentially the same, said capture zone channel containing at least one immobilized specific binding reagent, which immobilized specific binding reagent is capable of reacting and concentrating said labelled complex to form a detectable reaction product;

ii) applying a quantity of said fluid sample to said detection zone;

iii) permitting the fluid sample to flow from the detection zone from said plurality of different directions to the capture zone channel and any said analyte therein to form a detectable reaction product at said immobilized specific binding reagent in said capture zone channel; and

iv) identifying the presence of said at least one analyte in said fluid sample by detecting said detectable reaction product at said immobilized specific binding reagent in said capture zone channel.

38. The method of claim 37 wherein said at least one analyte is a cardiac analyte.

39. The method of claim 38 wherein three defined pathways are provided for identifying the presence of troponin I or troponin T together with identifying the presence of myoglobin and CK-MB.

40. The method of claim 39 wherein there is only one defined pathway provided for identifying the presence of troponin I or troponin T together with identifying the presence of myoglobin and CK-MB.

41. An analytical test device for determining the presence of at least one analyte in a fluid sample, said device comprising:

a dry porous carrier;

at least one detection zone covering at least a segment of an area of said carrier;

a sample circulation channel in operative communication with said detection zone, the sample being permitted to enter from said sample circulation channel into said detection zone from a plurality of different directions;

at least one capture zone channel having an entrance end which is in operative communication with the detection zone to permit sample to flow from the detection zone into the capture zone channel.

42. The device of claim 41 wherein said area is polygonal or forms part of a polygon.

43. The device of claim 42 wherein said area is rectangular of forms part of a rectangle.

44. An analytical test device suitable for determining the presence of at least one analyte contained in a liquid sample while permitting rapid and efficient flow of the sample through at least one defined pathway in which reactions determinative of the analyte take place, said device comprising:

a dry porous carrier through which a sample can flow by capillary action;

a sample delivery means through which the sample can be applied to the device and flow thereinto;

a sample circulation channel in operative communication with said sample delivery means, which is closed at its terminal ends and which circumscribes an area of the carrier;

at least one detection zone covering at least a segment of said area of the carrier, wherein at least a segment of the sample circulation channel conforms with a border of the detection zone and is in operative communication with said detection zone, across which border the sample is permitted to enter into the detection zone simultaneously from a plurality of different directions and to form a stream flowing away from said border, the detection zone containing at least one mobile, labelled specific binding reagent for the at least one analyte, which said at least one binding reagent is capable of reacting with said at least one analyte to form at least one labelled complex which is capable of moving along with said stream; and

at least one capture zone channel having an entrance end and a closed terminal end, the entrance end being in operative communication with the detection zone to permit said stream to flow from the detection zone into the capture zone channel, the capture zone channel containing at least one immobilized specific binding reagent, which at least one binding reagent is capable of reacting with and concentrating said at least one labelled complex to form at least one detectable reaction product.

45. The device of claim 44 wherein said sample circulation channel is polygonal or forms part of a polygon.

46. The device of claim 45 wherein said sample circulation channel is rectangular or forms part of a rectangle.

47. The device of claim 46 wherein the analyte is selected from troponin I, troponin T, myoglobin, CK-MB and mixtures thereof.

48. The device of claim 46 wherein the fluid sample is selected from whole blood, plasma, serum and urine.

49. The device of claim 46 wherein the dry porous carrier is nitrocellulose.

50. The device of claim 46 comprising two or more defined pathways, each pathway comprising a detection zone arranged on said area of the carrier, and a capture zone channel in operative communication with said detection zone.

51. The device of claim 46 wherein the label is a gold label.

52. The device of claim 46 wherein, in the detection zone, there is a mixture of a mobile labeled antibody which will react with one epitope on the analyte and a mobile biotin labelled antibody which will react with another epitope on the analyte, and the immobilized reagent is selected from the group consisting of streptavidin and avidin.

53. The device of claim 46 containing two defined pathways, one for determining the presence of myoglobin, the other for determining the presence of CK-MB.

54. The device of claim 46 containing three defined pathways, the first pathway for determining the presence of troponin I or troponin T, the second pathway for determining the presence of myoglobin and the third pathway for determining the presence of CK-MB.

55. The device of claim 46 wherein said at least one analyte is a cardiac analyte and said liquid sample is whole blood containing red blood cells and plasma, the dry porous carrier causing said red blood cells to separate chromatographically from the plasma to provide a plasma front moving in the capture zone channel and a red blood cell front upstream thereof, a detectable portion of the labelled complex being between the red blood cell front and the plasma front so that the labelled complex is substantially free of red blood cells when said labeled complex is captured in the capture channel.

56. The device of claim 55 wherein the mobile reagent in the detection zone is a labelled antibody which will react with one epitope on the analyte and the immobilized reagent is an antibody which will react with another epitope on the analyte.

57. The device of claim 55 wherein, in the detection zone, there is a mixture of a mobile labelled antibody which will react with one epitope on the analyte and a mobile biotin labelled antibody which will react with another epitope on the analyte, and the immobilized reagent is an avidin.

58. The device of claim 55 containing one defined pathway for determining the presence of troponin I or troponin T.

59. The device of claim 55 containing two defined pathways, one for determining the presence of myoglobin, the other for determining the presence of CK-MB.

60. The device of claim 55 containing three defined pathways, the first pathway for determining the presence of troponin I or troponin T, the second pathway for determining the presence of myoglobin and the third pathway for determining the presence of CK-MB.

61. The device of claim 60 containing one defined pathway for determining the presence of troponin I.

62. The device of claim 55 containing one defined pathway for determining the presence of troponin I or troponin T, the presence of myoglobin and the presence of CK-MB.

63. The device of claim 62 containing one defined pathway for determining the presence of troponin I, the presence of myoglobin and the presence of CK-MB.

64. An analytical test device suitable for determining the presence of at least one analyte contained in a liquid sample while permitting rapid and efficient flow of the sample through at least one defined pathway in which reactions determinative of the analyte take place, said device comprising:

a dry porous carrier through which a sample can flow by capillary action;

a sample delivery means through which the sample can be applied to the device and flow thereinto;

a sample circulation channel in operative communication with said sample delivery means, which is closed at its terminal ends and which circumscribes an area of the carrier, said sample circulation channel being polygonal or forming part of a polygon;

at least one detection zone covering at least a segment of said area of the carrier, wherein at least a segment of the sample circulation channel conforms with a border of the detection zone and is in operative communication with said detection zone, across which border the sample is permitted to enter into the detection zone simultaneously from a plurality of different directions and to form a stream flowing away from said border, the detection zone containing at least one mobile, labelled specific binding reagent for the at least one analyte, which said at least one binding reagent is capable of reacting with said at least one analyte to form at least one labelled complex which is capable of moving along with said stream; and

at least one capture zone channel having an entrance end and a closed terminal end, the entrance end being in operative communication with the detection zone to permit said stream to flow from the detection zone into the capture zone channel, the capture zone channel containing at least one immobilized specific binding reagent, which at least one binding reagent is capable of reacting with and concentrating said at least one labelled complex to form at least one detectable reaction product.

65. The device of claim 64 wherein said sample circulation channel is rectangular or forms part of a rectangle.

66. The device of claim 64 containing one defined pathway for determining the presence of troponin I, the presence of myoglobin and the presence of CK-MB.

67. The device of claim 64 wherein the dry porous carrier is nitrocellulose.

68. The device of claim 64 wherein the fluid sample is whole blood.

69. The device of claim 64 wherein, in the detection zone, there is a mixture of a mobile labeled antibody which will react with one epitope on the analyte and a mobile biotin labelled antibody which will react with another epitope on the analyte, and the immobilized reagent is selected from the group consisting of streptavidin and avidin.

70. An analytical test device suitable for determining the presence of at least one analyte contained in a liquid sample while permitting rapid and efficient flow of the sample through at least one defined pathway in which reactions determinative of the analyte take place, said device comprising:

a dry porous carrier through which a sample can flow by capillary action;

a sample delivery means through which the sample can be applied to the device and flow thereinto;

a sample circulation channel in operative communication with said sample delivery means, which is closed at its terminal ends and which circumscribes an area of the carrier, said sample circulation channel being rectangular or forming part of a rectangle;

at least one detection zone covering at least a segment of said area of the carrier, wherein at least a segment of the sample circulation channel conforms with a border of the detection zone and is in operative communication with said detection zone, across which border the sample is permitted to enter into the detection zone simultaneously from a plurality of different directions and to form a stream flowing away from said border, the detection zone containing at least one mobile, labelled specific binding reagent for the at least one analyte, which said at least one binding reagent is capable of reacting with said at least one analyte to form at least one labelled complex which is capable of moving along with said stream; and

at least one capture zone channel having an entrance end and a closed terminal end, the entrance end being in operative communication with the detection zone to permit said stream to flow from the detection zone into the capture zone channel, the capture zone channel containing at least one immobilized specific binding reagent, which at least one binding reagent is capable of reacting with and concentrating said at least one labelled complex to form at least one detectable reaction product.