Hemoglobin E Detection Using PCR with Confronting Two-Pair Primers
Sorasak Intorasoot PhD*, Rachanu Thongpung MSc*, Khajornsak Tragoolpua MSc*, Mongkol Chottayaporn MSc**
Affiliation : * Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai ** Division of Clinical Microscopy, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai
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).
Material and Method: 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.
Conclusion : Successful Hb E genotyping by PCR-CTPP was introduced. It allows for confirming and simulta-
neously detection with other thalassemia mutations.
Keywords : Hemoglobinopathy, Hemoglobin E, Beta-thalassemia, Polymerase chain reaction with confronting two-pair primers
