Patent ID: 6046019
Filing Date: 2000-04-04
Classification: C12Q,G01N,Y10T

Abstract:
A method of ascertaining the presence of granulocytes in a sample of whole blood which comprises the steps of:providing an assay medium which includes a diagnostic system consisting essentially of a peroxide and an oxidation-sensitive indicator which is capable of undergoing a myeloperoxidase-catalyzed color change reaction with said peroxide;withdrawing the blood sample from a subject;disrupting red blood cells in the sample by lysis and removing the debris of the disrupted red blood cells by trapping the granulocytes on a membrane and passing said debris through the membrane to isolate said granulocytes and to remove from the blood sample components in the sample that might interfere with a determination of the granulocytes in the sample;lysing the isolated granulocytes in the blood sample on said membrane to release intracellular myeloperoxidase present in said granulocytes;bringing the peroxide and indicator in the assay medium into contact with the released myeloperoxidase in that area of the membrane where the myeloperoxidase is released to catalyze color developing reactions between the peroxide and the oxidation-sensitive indicator; andemploying a change in the color of the indicator as evidence of the presence of granulocytes in the blood sample.