Methods and apparatus for blood typing with optical bio-discs

• US 20030224457A1
• Filed: 11/15/2002
• Published: 12/04/2003
• Est. Priority Date: 11/17/2000
• Status: Active Grant

• Current Assignee: Vindur Technologies, Inc.
• Original Assignee: Nagaoka & Co., Ltd.
• Inventors: Hurt, Susan Newcomb, McIntyre, Kevin Robert, Gordon, John Francis
• Granted Patent: US 7,026,131 B2
• Time in Patent Office: Days
• US Class Current: 435/7.2
• CPC Class Codes: G01N 33/80 involving blood groups or blood types or red blood cells white blood cells G01N33/56972
• CPC Class Codes: Y10T 436/106664 Blood serum or blood plasma standard or control

Abstract

This invention relates to clinical diagnostic assays, related optical bio-discs, and a disc-reading apparatus. The invention is directed to methods and apparatuses for performing immunohematology assays using an optical bio-disc analysis system. The invention is further directed to an optical bio-disc for performing an immunohematologic assay including a substrate having encoded information associated therewith. The encoded information may be readable by a disc drive assembly to control rotation of the disc. The disc may also include at least one target zone or capture zone associated with the substrate. The target zone is disposed at a predetermined location relative to a center of the substrate. The disc further includes a plurality of capture antibodies immobilized within the target zone, a flow channel, fluidic circuit, or analysis chamber associated with the target zone, and an input site in fluid communication with the analysis chamber.

Claims

1. A method for determining a blood type of an individual by direct typing on an optical bio-disc comprising:

applying red blood cells to at least one chamber in the optical bio-disc, the chamber surface including at least one capture field including a capture antibody, at least one positive control field, and at least one negative control field;

incubating the samples in the disc to promote antigen-antibody interaction;

placing the disc into an optical reader that supports it on a first side;

rotating the disc about an axis substantially perpendicular to the first side to separate non-captured cells from captured cells located on the chamber surface;

obtaining a measurement for the test field, the positive control field, and the negative control field; and

analyzing the measurement of the test field, the positive control field and the negative control field to determine blood type of the individual.

2. A method for determining the presence of antibodies to an ABO blood group of an individual'"'"'s blood sample by reverse-typing on an optical bio-disc including:

purifying serum from a blood sample;

creating a test sample by mixing the serum with cells of a known ABO blood group;

injecting the test sample into at least one channel in the optical bio-disc, thereby delivering the test sample onto a capture field including a cell binding molecule;

incubating the test sample on the capture field to allow the agglutinated and non-agglutinated cells to bind to the cell binding molecule;

placing the disc into an optical reader that supports it on a first side;

rotating the disc about an axis substantially perpendicular to the first side;

scanning the chamber with an incident beam of electromagnetic radiation by rotating the disc about an axis substantially perpendicular to the first side by moving the incident beam in a direction radial to the axis;

detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc;

converting the return beam into an output signal;

analyzing the output signal to determine the presence of agglutinated cells bound on the capture field; and

determining the presence of antibodies in the serum.

3. A method for determining the presence of antibodies to an ABO blood group of an individual'"'"'s blood sample by reverse-typing on an optical bio-disc comprising:

applying a blood sample to at least one microfluidic channel in the optical bio-disc including a separation chamber with at least one microfilter, at least one mixing chamber, and at least one capture chamber;

spinning the optical bio-disc for a first time at a first speed to effect separation of the blood sample into cells and serum in the separation chamber;

spinning the optical bio-disc for a second time at a second speed higher than the first, the second speed effecting movement of the serum through the microfluidic channel into a mixing chamber;

adding cells of a known ABO blood group cells into the mixing chamber containing serum;

spinning the optical bio-disc for a third time in one direction and alternately in another direction at least once to effect mixing of the serum and the cells;

incubating the cells in the serum for a sufficient period of time to allow antibody-antigen binding;

spinning the optical bio-disc for a fourth time at a third speed higher than the second, the third speed effecting movement of the cells into a capture chamber, the capture chamber including surface with a molecule that binds cells;

incubating the sample in the capture chamber to promote cell binding to the chamber surface;

spinning the disc for a fifth time to remove non-bound cells from the capture field;

scanning the chamber with an incident beam of electromagnetic radiation by rotating the disc about an axis substantially perpendicular to the first side by moving the incident beam in a direction radial to the axis;

detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc;

converting the return beam into an output signal;

analyzing the output signal to determine the presence of agglutinated cells; and

determining the presence of antibodies in the sample.

4. A method for determining the presence of antibodies to a blood group type in an individual by antibody-typing on an optical bio-disc comprising:

purifying serum from a blood sample;

creating at least one sample by mixing serum with cells of a known blood group phenotype;

injecting at least one sample into at least one channel in the optical bio-disc, thereby delivering the sample onto a capture field including a cell binding molecule;

placing the disc into an optical reader that supports it on a first side;

rotating the disc about an axis substantially perpendicular to the first side;

scanning the chamber with an incident beam of electromagnetic radiation by rotating the disc about an axis substantially perpendicular to the first side by moving the incident beam in a direction radial to the axis;

detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc;

converting the return beam into an output signal;

analyzing the output signal to determine the presence of cells bound to the capture field; and

determining the presence of blood group antibodies.

5. A method for determining the presence of antibodies to a blood group type of an individual by antibody-typing on an optical bio-disc comprising:

applying a blood sample to at least one microfluidic channel in the optical bio-disc including a separation chamber with at least one microfilter, at least one mixing chamber, and at least one capture chamber;

spinning the optical bio-disc for a first time at a first speed to effect separation of the blood sample into cells and serum in the separation chamber;

spinning the optical bio-disc for a second time at a second speed higher than the first, the second speed effecting movement of the serum through the microfluidic channel into a mixing chamber;

adding cells of a known blood group phenotype into the mixing chamber containing serum;

spinning the optical bio-disc for a third time in one direction and alternately in another direction at least once to effect mixing of the serum and the cells of a known blood group phenotype;

incubating the cells of a known blood group phenotype in the serum for a sufficient period of time to allow antibody-antigen binding;

spinning the optical bio-disc for a fourth time at a third speed higher than the second, the third speed effecting movement of the cells of a known blood group phenotype into of a capture chamber, the capture chamber including a surface with an anti-human immunoglobulin molecule;

incubating the sample in the capture chamber to promote binding of the cells of a known blood group phenotype to the chamber surface;

spinning the optical bio-disc for a fifth time to remove non-bound cells of a known blood group phenotype;

scanning the chamber with an incident beam of electromagnetic radiation by rotating the disc about an axis substantially perpendicular to the first side by moving the incident beam in a direction radial to the axis;

detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc;

converting the return beam into an output signal;

analyzing the output signal to determine if the cells of a known blood group phenotype are captured; and

determining the presence of blood group antibodies.

6. An apparatus for determining a blood group type of an individual comprising:

an optical bio-disc including at least one capture chamber including;

a layer including a first capture antibody, and

a layer including a second capture antibody bound by the first capture antibody, the second capture antibody being specific for a blood group antigen;

a disc drive assembly;

an optical reader; and

software for blood group analysis.

7. An optical-bio disc for performing a blood-typing assay, said disc comprising:

a substrate;

a separation chamber associated with said substrate, said separation chamber including a first inlet port;

a filter means associated with said separation chamber;

a first mixing chamber in fluid communication with said separation chamber, said first mixing chamber including a second inlet port;

a second mixing chamber in fluid communication with said separation chamber, said second mixing chamber including a third inlet port;

a first analysis chamber in fluid communication with said first mixing chamber, said first analysis chamber including a capture field; and

a second analysis chamber in fluid communication with said second mixing chamber, said second analysis chamber including a capture field.

8. A method for determining a blood group type of an individual by direct typing on an optical bio-disc, said method comprising:

loading reagent antibodies into at least one chamber in the optical bio-disc, said at least one chamber having a bio-matrix packed therein such that the packing density allows single cells to pass through and prevents agglutinated cells from entering;

loading red blood cells to said at least one chamber in the optical bio-disc to create an assay solution including said reagent antibodies and said red blood cells;

incubating said assay solution in the disc to promote antigen-antibody interaction and red blood cell agglutination;

placing the disc into an optical reader to support the disc therein;

rotating the disc about an axis substantially perpendicular to a substrate layer of the disc to thereby direct said assay solution through said bio-matrix to thereby separate non-agglutinated red blood cells from agglutinated red blood cells;

directing a beam of electromagnetic radiation to scan said at least one chamber; and

analyzing the return signal from the beam to determine the amount of red blood cell agglutination to determine the blood group of the individual.

9. A method for determining the presence of antibodies to an ABO blood group of an individual'"'"'s blood sample by reverse-typing on an optical bio-disc, said method comprising:

loading a blood sample into at least one microfluidic channel in the optical bio-disc including a preparation chamber with at least one microfilter, at least one mixing chamber, and at least one separation chamber, said separation chamber having a bio-matrix packed therein such that the packing density allows single cells to pass through and prevents agglutinated cells from entering; and

spinning the disc for a first time at a first speed to effect separation of the blood sample into cells and serum in the preparation chamber.

10. The method according to claim 9 including the further step of spinning the disc for a second time at a second speed greater than the first, the second speed effecting movement of the serum through the microfluidic channel into the mixing chamber.

11. The method according to claim 10 including the further step of adding cells of a known ABO blood group into the mixing chamber containing the serum.

12. The method according to claim 11 including the further step of spinning the disc for a third time at a third speed in one direction and alternately in another direction at least once to effect mixing of the serum and the cells of a known ABO blood group.

13. The method according to claim 12 including the further step of incubating the cells of a known ABO blood group with the serum for a sufficient period of time to allow antibody-antigen binding and cell agglutination.

14. The method according to claim 13 including the further step of spinning the disc for a fourth time at a fourth speed greater than the second, the fourth speed effecting movement of the cells of a known ABO blood group into the separation chamber and into the bio-matrix so that agglutinated cells are separated from non-agglutinated cells.

15. The method according to claim 14 including the further step of scanning the chamber with an incident beam of electromagnetic radiation by rotating the disc about an axis substantially perpendicular to a substrate portion of the disc and by moving the incident beam in a direction radial to the axis.

16. The method according to claim 15 including the further step of detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc.

17. The method according to claim 16 including the further step of converting the return beam into an output signal.

18. The method according to claim 17 including the further step of analyzing the output signal to determine the presence of agglutinated cells to thereby determine the presence of antibodies in the sample.

19. A method for determining the presence of antibodies of a blood group type in an individual by antibody-typing on an optical bio-disc, said method comprising:

purifying serum from a blood sample;

creating at least one assay solution by mixing the purified serum with cells of a known blood group phenotype;

loading the assay solution into at least one channel in the optical bio-disc, said channel containing a bio-matrix packed therein such that the packing density allows single cells to pass through and prevents agglutinated cells from entering;

incubating the assay solution for a sufficient period of time to allow antibody-antigen binding and agglutination;

placing the disc into an optical reader to support the disc therein;

rotating the disc about an axis substantially perpendicular to a substrate layer of the disc to direct the assay solution through the bio-matrix so that the non-agglutinated cells are separated from the agglutinated cells;

scanning the chamber with an incident beam of electromagnetic radiation by rotating the disc about said axis and by moving the incident beam in a direction radial to the axis;

detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc;

converting the return beam into an output signal; and

analyzing the output signal to determine the presence and extent of agglutination to thereby determine the presence of blood group antibodies.

20. An optical-bio disc for performing an immunohematologic assay, said disc comprising:

a substrate having encoded information associated therewith, said encoded information being readable by a disc drive assembly to control rotation of the disc;

a separation chamber associated with said substrate, said separation chamber including a first inlet port;

filter means associated with said separation chamber;

a first mixing chamber in fluid communication with said separation chamber, said first mixing chamber including a second inlet port;

a second mixing chamber in fluid communication with said separation chamber, said second mixing chamber including a third inlet port;

a first analysis chamber in fluid communication with said first mixing chamber, said first analysis chamber including a bio-matrix; and

a second analysis chamber in fluid communication with said second mixing chamber, said second analysis chamber including a bio-matrix.

21. An optical bio-disc for performing an immunohematologic assay, said bio-disc comprising:

a substrate having encoded information associated therewith, said encoded information being readable by a disc drive assembly to control rotation of the disc;

at least one capture field associated with said substrate, said capture field disposed at a predetermined location relative to a center of said substrate;

a plurality of capture antibodies immobilized within said capture field;

a flow channel associated with said capture field; and

an input site in fluid communication with said flow channel.

22. The optical bio-disc according to claim 21 wherein said input site is implemented to receive an assay solution, said assay solution including buffer, type A reagent cells, type B reagent cells, and a serum sample, such that when said assay solution is deposited in said flow channel, said type A and type B reagent cells move into said capture field and binding occurs between the surface antigens on the type A or type B cells and their respective capture antibodies to thereby place said type A and type B cells in their respective capture fields.

23. The optical bio-disc according to claim 22 wherein when a beam of electromagnetic radiation is directed into the capture field, whereby the presence and amount of agglutination may be determined from the signal returned from said beam.

24. A method for determining the presence of antibodies of an ABO blood group of an individual'"'"'s blood sample by reverse-typing on an optical bio-disc, said method comprising:

loading a blood sample to at least one microfluidic channel in the optical bio-disc including a separation chamber with at least one microfilter, at least one mixing chamber, and at least one capture chamber; and

spinning the disc a first time at a first speed to effect separation of the blood sample into blood cells and serum in the separation chamber.

25. The method according to claim 24 including the further step of spinning the disc a second time at a second speed greater than the first, the second speed effecting movement of the serum through the microfluidic channel into a mixing chamber.

26. The method according to claim 25 including the further step of adding type A and type B reagent cells into the mixing chamber containing the serum.

27. The method according to claim 26 including the further step of spinning the disc a third time at a third speed in one direction and alternately in another direction at least once to effect mixing of the serum and the reagent cells.

28. The method according to claim 27 including the further step of incubating the reagent cells in the serum for a sufficient period of time to allow antibody-antigen binding and cell agglutination.

29. The method according to claim 28 including the further step of spinning the optical bio-disc for a fourth time at a fourth speed greater than the second, the fourth speed effecting movement of the reagent cells into said capture chamber, said capture chamber including a surface with a capture agent.

30. The method according to claim 29 wherein said capture agent is selected from the group comprising an antibody directed against type A reagent cells and an antibody directed against type B reagent cells.

31. The method according to claim 30 including the further step of incubating the sample in the capture chamber to promote cell binding to the capture agent.

32. The method according to claim 31 including the further step of spinning the disc for a fifth time to remove non-bound reagent cells from the capture field.

33. The method according to claim 32 including the further step of scanning the chamber with an incident beam of electromagnetic radiation by rotating the disc about an axis substantially perpendicular to said substrate and by moving the incident beam in a direction radial to the axis.

34. The method according to claim 33 including the further step of detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc.

35. The method according to claim 34 including the further step of converting the return beam into an output signal.

36. The method according to claim 35 including the further step of analyzing the output signal to determine the presence and amount of agglutinated and non-agglutinated reagent cells to thereby determine the presence of antibodies in the sample.

37. A method for determining the presence of antibodies of a blood group type in an individual by antibody-typing on an optical bio-disc, said method comprising:

purifying serum from a blood sample;

creating at least one assay solution by mixing serum with reagent cells expressing antigens of any minor blood group, and anti-human globulin;

loading the assay solution into at least one channel in the optical bio-disc, thereby delivering the assay solution onto a capture field including at least one capture antibody;

incubating the cells in the serum for a sufficient period of time to allow antibody-antigen binding and agglutination;

incubating the sample on the capture field to allow the cells to bind to their respective capture antibodies in the capture field;

spinning the disc to separate unbound cells from cells bound in the capture fields;

placing the disc into an optical reader;

rotating the disc about an axis substantially perpendicular to a substrate layer of the disc;

scanning the chamber with an incident beam of electromagnetic radiation by rotating the disc about said axis and by moving the incident beam in a direction radial to the axis;

detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc;

converting the return beam into an output signal; and

analyzing the output signal to determine the presence and amount of agglutination to thereby determine the presence of blood group antibodies.

38. An optical bio-disc for performing an immunohematologic assay, said bio-disc comprising:

a substantially circular substrate having an center and an outer edge;

a channel layer having a fluidic circuit formed therein, said channel layer being associated with said substrate;

a cap portion associated with said channel layer;

at least one capture field associated with said substrate, said capture field disposed at a predetermined location within said fluidic circuit;

a plurality of capture antibodies immobilized within said capture field.

39. The optical bio-disc according to claim 38 further comprising a semi-reflective layer associated with said substrate.

40. The optical bio-disc according to claim 39 wherein said substrate includes encoded information associated therewith, said encoded information being readable by a disc drive assembly to control rotation of the disc.

41. The optical bio-disc according to claim 40 wherein said fluidic circuit includes an inlet port, an analysis chamber, and a vent port.

42. The optical bio-disc according to claim 41 wherein said capture field is located within said analysis chamber.

43. The optical bio-disc according to claim 42 wherein said analysis chamber is substantially directed along an arc segment of an annular ring within said center and outer edge of said substrate.

44. The optical bio-disc according to claim 42 wherein said analysis chamber is substantially directed along a pre-determined radius on the disc.

45. The optical bio-disc according to either claim 43 or 44 wherein said inlet port is implemented to receive an assay solution, said assay solution including buffer, type A reagent cells, type B reagent cells, and a serum sample, such that when said assay solution is deposited in said fluidic circuit, said type A and type B reagent cells move into said analysis chamber and into said capture field and binding occurs between the surface antigens on the type A or type B cells and their respective capture antibodies to thereby place said type A and type B cells in their respective capture fields.

46. The optical bio-disc according to claim 45 wherein when a beam of electromagnetic radiation is directed into the capture field, the presence and amount of reagent cell agglutination is determined from the signal returned from said beam.

47. A method of making an optical bio-disc for immunohematologic testing, said method of making comprising the steps of:

providing a substrate having a center and an outer edge;

encoding information on an information layer associated with the substrate, said encoded information being readable by a disc drive assembly to control rotation of the disc;

forming a capture field in association with said substrate, said capture field disposed at a predetermined location relative to said center of said substrate;

depositing within said capture field, one more capture agents;

forming an analysis chamber in fluid communication with said capture field;

designating an input site associated with the analysis chamber, the input site implemented to receive a sample to be tested for blood type, so that when the sample is deposited in the analysis chamber and the disc is rotated, the sample moves into the capture field and binding occurs between an analyte in the sample and the one or more capture agents to thereby place the analyte in the capture field.

48. The method according to claim 47 wherein said analyte is selected from the group comprising red blood cells and typed reagent cells.

49. The method according to claim 48 wherein said analysis chamber is substantially directed along an arc segment of an annular ring within said center and outer edge of said substrate.

50. The optical bio-disc made according to claim 49.

51. A method of using the optical bio-disc made according to claim 49, said method of using comprising:

purifying serum from a blood sample;

creating at least one assay solution by mixing serum with typed reagent cells expressing antigens of any minor blood group, and anti-human globulin;

loading the assay solution into the analysis chamber in the optical bio-disc, thereby delivering the assay solution onto the capture field;

incubating the typed reagent cells in the serum for a sufficient period of time to allow antibody-antigen binding and cell agglutination;

incubating the sample on the capture field to allow the cells to bind to the one or more capture agents in the capture field;

spinning the disc to separate unbound typed reagent cells from typed reagent cells bound in the capture fields;

placing the disc into an optical reader;

rotating the disc about an axis substantially perpendicular to the substrate;

scanning the analysis chamber with an incident beam of electromagnetic radiation by rotating the disc about said axis and by moving the incident beam in a direction radial to the axis;

detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc;

converting the return beam into an output signal; and

analyzing the output signal to determine the presence and amount of agglutination to thereby determine the presence of blood group antibodies.

52. A method of using the optical bio-disc made according to claim 49, said method of using comprising:

applying a blood sample into the analysis chamber;

incubating the blood sample in the analysis chamber for a sufficient time to allow binding of red blood cells in the blood sample to the one or more capture agents in the capture zone;

placing the disc into an optical disc reader;

rotating the disc using the optical disc reader about an axis substantially perpendicular to the substrate at a pre-determined speed and time to separate non-bound cells from bound cells on the capture field;

scanning the analysis chamber with an incident beam of electromagnetic radiation by rotating the disc about said axis and by moving the incident beam in a direction radial to the axis;

detecting a return beam of electromagnetic radiation formed by at least a part of the incident beam after interacting with the disc;

converting the return beam into an output signal; and

analyzing the output signal to determine the presence blood cells in the capture field to thereby determine the presence of blood group of said blood sample.

53. The method according to either claim 51 or 52 wherein said capture agent is selected from the group comprising lectin, a monoclonal antibody, a polyclonal antibody, and an oligonucleotide.