Hemoglobin E detection using PCR with confronting two-pair primers.
OBJECTIVE
To develop and apply the polymerase chain reaction with confronting two-pair primers (PCR-CTPP) for detection and identification of hemoglobin E (Hb E).


METHODS
Fifty unrelated northern Thais were included in the present study. DNA was extracted from peripheral blood mononuclear cells and targeted to amplify by PCR-CTPP. The amplified product was analyzed and compared with the reference hemoglobin electrophoresis and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis.


RESULTS
The results validated a completely concordant among these three methods consisting of 74%, 24%, and 2% identified as normal, heterozygous, and homozygous Hb E type, respectively.


CONCLUSIONS
Successful Hb E genotyping by PCR-CTPP was introduced. It allows for confirming and simultaneously detection with other thalassemia mutations.
Intorasoot
Sorasak
S
Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, 110 Indrawaroros Rd, Sripoom, Mueang, Chiang Mai 50200, Thailand. isorasak@hotmail.com
Thongpung
Rachanu
R
Tragoolpua
Khajornsak
K
Chottayaporn
Mongkol
M
D004247:DNA; D017931:DNA Primers; D005784:Gene Amplification; D005838:Genotype; D006446:Hemoglobin E / Q000032:analysis* / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics*; D013785:Thailand; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics
D017931:DNA Primers; D004247:DNA; D006446:Hemoglobin E