Allele-specific amplification for the identification of several hemoglobin variants.
Eight known abnormal hemoglobins in more than 40 patients have been identified with an allele-specific amplification procedure. These are mutants of the A gamma (Hb F-Sardinia), the delta (Hb B2), the alpha (Hb G-Philadelphia), and the beta (Hbs S, C, E, D-Los Angeles, O-Arab) globin genes. Adjustment to an unusually high annealing temperature improves the procedure greatly, allowing excellent discrimination between normal and mutant alleles, irrespective of the particular mismatch. The discrimination includes the Hbs C and E in the same reaction mixture-multiplex allele-specific amplification. The procedure can successively use primers with 3'-penultimate critical nucleotides, degenerative ends, and potentially dimerizing properties.
Postnikov YuV
Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta 30912-2100.
Molchanova
T P
TP
Huisman
T H
TH
D000483:Alleles; D001482:Base Composition; D001483:Base Sequence; D016002:Discriminant Analysis; D005784:Gene Amplification; D005820:Genetic Testing; D005838:Genotype; D005914:Globins / Q000235:genetics*; D006451:Hemoglobin, Sickle; D006454:Hemoglobins / Q000235:genetics*; D006455:Hemoglobins, Abnormal / Q000235:genetics; D006579:Heterozygote; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation
D006451:Hemoglobin, Sickle; D006454:Hemoglobins; D006455:Hemoglobins, Abnormal; D005914:Globins; C026237:hemoglobin D Punjab; C032031:hemoglobin Arab