title: Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays.
abstract: Beta-thalassemia is a life-threatening inherited blood disorder. Rapid characterization of β-globin gene mutations is necessary because of the high frequency of Malaysian β-thalassemia carriers. A combination real-time polymerase chain reaction genotyping assay using TaqMan probes was developed to confirm β-globin gene mutations. In this study, primers and probes were designed to specifically identify 8 common β-thalassemia mutations in the Malaysian Malay and Chinese ethnic groups using the Primer Express software. "Blind tests" using DNA samples from healthy individuals and β-thalassemia patients with different genotypes were performed to determine the specificity and sensitivity of this newly designed assay. Our results showed 100% sensitivity and specificity for this novel assay. In conclusion, the TaqMan genotyping assay is a straightforward assay that allows detection of β-globin gene mutations in less than 40 min. The simplicity and reproducibility of the TaqMan genotyping assay permit its use in laboratories as a rapid and cost-effective diagnostic tool for confirmation of common β-thalassemia mutations in Malaysia.
authors.affiliation: Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: S L
authors.initials: SL
authors.lastname: Kho
authors.forename: K H
authors.initials: KH
authors.lastname: Chua
authors.forename: E
authors.initials: E
authors.lastname: George
authors.forename: J A M A
authors.initials: JA
authors.lastname: Tan
mesh_terms: D044466:Asian People / Q000208:ethnology / Q000235:genetics*; D016022:Case-Control Studies; D060005:Genotyping Techniques / Q000379:methods*; D006242:Haptoglobins / Q000235:genetics; D006801:Humans; D008296:Malaysia; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D016037:Single-Blind Method; D017086:beta-Thalassemia / Q000208:ethnology / Q000235:genetics*
chemical_list: C485359:HP protein, human; D006242:Haptoglobins