title: Rapid identification of heterozygous and homozygous hemoglobin constant spring by SYTO9 with a high resolution melting analysis.
abstract: BACKGROUND
  Gel-electrophoresis and ethidium bromide are not ideally suited to large scale analysis in clinical laboratories.
  
  
  METHODS
  Amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) specific for Hb CS was performed in 10 blood samples from normal individuals and 61 samples containing a peak of Hb CS when analyzed by capillary electrophoresis. Heterozygosity of Hb CS was identified using SYTO9 and high resolution melting (HRM) analysis method.
  
  
  RESULTS
  Specific peak heights of amplified fragments of wild type and Hb CS alleles were observed in the heterozygote. Only one peak height of amplified fragments of the wild type allele was observed in the normal individual while only one peak height of amplified fragments of Hb CS allele was observed in the homozygote. HRM analysis interpretation results were completely consistent with the interpretation results from gel-electrophoresis.
  
  
  CONCLUSIONS
  SYTO9 HRM analysis may be used as an alternative for rapid diagnosis of heterozygosity of Hb CS.
authors.affiliation: Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang-Mai University, Chiang-Mai, Thailand. sakornmi001@gmail.com
authors.forename: Sakorn
authors.initials: S
authors.lastname: Pornprasert
authors.forename: Panyasai
authors.initials: P
authors.lastname: Sitthichai
authors.forename: Jarurin
authors.initials: J
authors.lastname: Waneesorn
authors.forename: Kanyakan
authors.initials: K
authors.lastname: Kongthai
authors.forename: Panthong
authors.initials: P
authors.lastname: Singboottra
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D006454:Hemoglobins / Q000235:genetics*; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D006454:Hemoglobins