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Detection of hemoglobin E heterozygotes by using polymerase chain reaction and direct DNA sequencing: report of a
1Department of Internal Medicine, Kaoshiung Medical College, Taiwan, R.O.C.
Abstract:
From an overseas Chinese born in Thailand, an extraordinarily high level of A2 + E band (20.2%) was found during routine cellulose acetate Hb electrophoresis. With a suspicion of Hb E, the beta-globin gene was studied by using the polymerase chain reaction (PCR) and direct DNA sequencing method. A 1.4 kb fragment covering the whole beta-globin gene was amplified by PCR. Sequence analysis of the amplified product revealed a GAG > AAG transversion at codon 26, which resulted in an amino acid substitution of lysine for glutamic acid. A normal sequence at the corresponding codon was noted in the other allele; hence, this patient is a heterozygote of Hb E. PCR and direct DNA sequencing provide a rapid method for detection of Hb E from a small amount of DNA.