Microfluidic device and leucocyte antigen mediated microfluidic assay

摘要

Provided is a leucocyte antigen mediated microfluidic assay for testing a leucocyte containing fluid from a subject for prior exposure to a predetermined antigen which would have caused the leucocyte to become presensitized to the antigen comprising: performing a first test comprising the steps of: a) placing a microvolume first sample of the subjects leucocyte containing fluid in a first sample microchamber; b) transporting the first sample to a first reaction microchamber containing the predetermined antigen and an antigen accelerator and allowing them to react together for a predetermined time; c) transporting the reacted first sample to a first observation microchamber; d) mixing one or more leucocyte observation colorants with the reacted first sample during transportation to the observation microchamber or in the observation microchamber; e) optically performing an optical scan of the reacted sample in the first observation microchamber under conditions which allow the measurement of at least 1 measurement factor of the leucocytes in the reacted first sample; performing a second test comprising the steps of: a) placing a microvolume second sample of the subjects leucocyte containing fluid in a second sample microchamber; b) transporting the second sample to an antigen free second reaction microchamber containing the same antigen accelerator as the first test and allowing them to react for the predetermined time; c) transporting the second sample to a second observation microchamber; d) mixing the same leucocyte observation colorants as the first test with the reacted second sample during transport to the second observation microchamber or in the second observation microchamber; e) optically performing an optical scan of the reacted second sample in the second observation microchamber under conditions that allow the measurement of the same leucocyte measurement factors measured in the first test; comparing the results of the first test scan to the second test scan to determine if any differences between the two scans are due to a leucocyte-antigen mediated response wherein the optical scan is accomplished by multi-wavelength fluorescence image analysis. Also provided is a corresponding device for the assay.