ALLELE-SPECIFIC AMPLIFICATION FOR THE IDENTIFICATION OF SEVERAL HEMOGLOBIN-VARIANTS

Eight known abnormal hemoglobins in more than 40 patients have been id entified with an allele-specific amplification procedure. These are mu tants of the A(gamma) (Hb F-Sardinia), the delta (Hb B2), the alpha (H b G-Philadelphia), and the beta (Hbs S, C, E, D-Los Angeles, O-Arab) g lobin genes. Adjustment to an unusually high annealing temperature imp roves the procedure greatly, allowing excellent discrimination between normal and mutant alleles, irrespective of the particular mismatch. T he discrimination includes the Hbs C and E in the same reaction mixtur e - multiplex allele-specific amplification. The procedure can success ively use primers with 3'-penultimate critical nucleotides, degenerati ve ends, and potentially dimerizing properties.