title: Diagnosis of alpha-thalassemia-1 Southeast Asian type deletion and fetal gender by single-tube multiplex real-time PCR.
abstract: BACKGROUND
  The combination of an x-linked hematologic disorder with a heterozygous alpha-thalassemia-1 Southeast Asian (SEA) type deletion might lead to severe anemia in male infants. This study is to develop a simple and cost-effective single tube multiplex real-time PCR for the diagnosis of alpha-thalassemia-1 SEA type deletion and detect fetal gender.
  
  
  METHODS
  Multiplex real-time polymerase chain reaction (PCR) for the detection of alpha-thalassemia-1 SEA type deletion gene, wild type alpha-globin gene, and sex-determining region Y (SRY) gene was validated by analysis of 95 cord blood samples (60 normal individuals, 28 heterozygous of alpha-thalassemia-1 SEA type deletion and 7 Bart's hydrops fetalis). The change in threshold cycle (deltaC(T)) was analyzed by subtracting the C(T-mutant) from C(T-wild) type.
  
  
  RESULTS
  Mean deltaC(T) values were significantly different among these three groups, and a SRY gene determination was 100% in concordance with fetal genders. Furthermore, analysis of fetal gender did not affect the deltaC(T) of alpha-thalassemia-1 SEA detection.
  
  
  CONCLUSIONS
  Combined alpha-thalassemia-1 SEA type detection and fetal gender determination in a single-tube multiplex real-time PCR is an alternative assay for a conventional method for the diagnosis of alpha-thalassemia-1 SEA deletion and fetal gender.
authors.affiliation: Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang-Mai University, Chiang-Mai, Thailand.
authors.forename: Sarinee
authors.initials: S
authors.lastname: Srithep
authors.forename: Kanyakan
authors.initials: K
authors.lastname: Kongthai
authors.forename: Sakorn
authors.initials: S
authors.lastname: Pornprasert
mesh_terms: D005260:Female; D017353:Gene Deletion; D024282:Genes, sry / Q000235:genetics; D006801:Humans; D008297:Male; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D017085:alpha-Thalassemia / Q000235:genetics*