Detection of hemoglobin E heterozygotes by using polymerase chain reaction and direct DNA sequencing: report of a case.
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.
Department of Internal Medicine, Kaoshiung Medical College, Taiwan, R.O.C.
T C
TC
Liu
C P
CP
Lin
P H
PH
Chen
L S
LS
Lee
J G
JG
Chang
D000328:Adult; D044466:Asian People / Q000235:genetics; D001483:Base Sequence; D002681:China / Q000208:ethnology; D004247:DNA / Q000032:analysis*; D005260:Female; D005914:Globins / Q000235:genetics; D006446:Hemoglobin E / Q000235:genetics*; D006579:Heterozygote; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D013785:Thailand
D005914:Globins; D004247:DNA; D006446:Hemoglobin E