Abstract
Glycophorin A (GPA) is the major glycoprotein on human erythrocyte membranes, and bears M and N blood group determinants. Genotoxic exposure causes erythroid precursor cells to mutate at the glycophorin. A locus, producing phenotypes of rare variant erythrocytes that lack the expression of one form of the protein. The BR6 assay is now the most widely used method to determine the variant frequency (Vf) by immunolabelling and flow cytometry. In this study, two kinds of labelling were used. Second antibody labelling, namely BRIC157-sheep anti-mouse IgG-FITC binding, increased the sensitivity, while the specificity and the precision decreased. NO/NN Vf of 10 normal samples conform with previous reports, 7.96 ± 3.05/2.24 ± 1.05 (x10-6) and 10.12 ± 3.2/3.95 ± 1.02 (x10-6) of direct and indirect dyeing respectively. Some factors affect the dyes emitting fluorescence, and the optimal temperature of immunolabelling is under 20(C.
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