title: [Improved hybridization probe method for genetic analysis of beta-thalassemia].
abstract: There are a number of methods for genetic diagnosis of a point mutation and/or deletion/insertion of several nucleotides. However, none of these methods are satisfactory with respect to sensitivity, accuracy and cost effectiveness. In 2003, the Hybridization Probe method was first reported, but it was unsatisfactory. We attempted to improve this method by separating the step of using real-time PCR with melting curve analysis. In our improved method, or Hybri Probe method, the sensitivity and accuracy of Tm were remarkably improved by employing H2O and Hi-Di Formamide. In order to detect several beta-thalassemia mutations, relevant probes of basically wild sequence were prepared, and optimal conditions for detecting mutations were studied. This method yielded highly sensitive and accurate results. This Hybri Probe method is available for SNPs and microsatellite polymorphisms, and it may also be useful for the detection of gene abnormalities other than those in beta-thalassemia.
authors: Faculty of Medicine and Health Science, Yamaguchi University, Ube 755-8505, Japan. yamasiro@yamaguchi-u.ac.jp
authors: Yasuhiro
authors: Y
authors: Yamashiro
authors: Yukio
authors: Y
authors: Hattori
authors: Minako
authors: M
authors: Hino
authors: Chiemi
authors: C
authors: Furuya
authors: Azusa
authors: A
authors: Fukamitu
authors: Yuri
authors: Y
authors: Suehisa
mesh_terms: D015342:DNA Probes; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D015336:Molecular Probe Techniques; D017354:Point Mutation; D012680:Sensitivity and Specificity; D044366:Transition Temperature; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics*
chemical_list: D015342:DNA Probes; D055544:beta-Globins