Methods and apparatus for blood typing with optical bio-discs
View Patent ↗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.
1. 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 capture chamber surface;
spinning the disc for a fifth time to remove non-bound cells from the capture chamber;
scanning the capture chamber with an incident beam of electromagnetic radiation by rotating the disc about an axis substantially perpendicular to a first face of the disc 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;
analyzing the output signal to determine the presence of agglutinated cells; and
determining the presence of antibodies in the sample.
2. 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 capture chamber surface;
spinning the optical bio-disc for a fifth time to remove non-bound cells of a known blood group phenotype;
scanning the capture chamber with an incident beam of electromagnetic radiation by rotating the disc about an axis substantially perpendicular to a first face of the disc 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;
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.
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, 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;
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;
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;
adding cells of a known ABO blood group into the mixing chamber containing the serum;
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;
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; and
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.
4. The method according to claim 3 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.
5. The method according to claim 4 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.
6. The method according to claim 5 including the further step of converting the return beam into an output signal.
7. The method according to claim 6 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.
8. 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;
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;
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;
adding type A and type B reagent cells into the mixing chamber containing the serum;
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;
incubating the reagent cells in the serum for a sufficient period of time to allow antibody-antigen binding and cell agglutination; and
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.
9. The method according to claim 8 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.
10. The method according to claim 9 including the further step of incubating the sample in the capture chamber to promote cell binding to the capture agent.
11. The method according to claim 10 including the further step of spinning the disc for a fifth time to remove non-bound reagent cells from the capture field.
12. The method according to claim 11 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.
13. The method according to claim 12 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.
14. The method according to claim 13 including the further step of converting the return beam into an output signal.
15. The method according to claim 14 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.