Hemoglobin E detection using PCR with confronting two-pair primers

J Med Assoc Thai. 2008 Nov;91(11):1677-80.

Abstract

Objectives: 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 simultaneously detection with other thalassemia mutations.

Publication types

  • Validation Study

MeSH terms

  • DNA
  • DNA Primers*
  • Gene Amplification
  • Genotype
  • Hemoglobin E / analysis*
  • Hemoglobin E / genetics
  • Humans
  • Polymerase Chain Reaction / instrumentation*
  • Polymerase Chain Reaction / methods
  • Polymorphism, Restriction Fragment Length / genetics*
  • Thailand
  • beta-Thalassemia / diagnosis*
  • beta-Thalassemia / genetics

Substances

  • DNA Primers
  • DNA
  • Hemoglobin E