title: [Simple and rapid analysis of beta-thalassemia mutation by sequence-specific amplification].
abstract: This study was done to detect and diagnose beta-thalassemia (beta-Thal) gene quickly. We applied sequence specific Amplification (SSA) method to the analysis. 13 kinds of beta-Thal and two kinds of hemoglobin variants were able to detect under the same PCR condition. These mutations were found frequently in ten countries of Asian region (the southern part of China, Vietnam, Cambodia, Thailand, Myanmar, Malaysia, Singapore, Indonesia, Pakistan, India), and 15 kinds in total (-28CapA-->G, CD5-CT, CD8/9+-G, CD15G-->A, CD17A-->T, IVSI-1G-->T, CD41/42-4del, CD16-C, CD26G-->A(betaE), IVSI-5G-->C, CD35C-->A, CD71/72 +A, CD6A-->T (betaS), -619del, IVSII-654C-->T). More than 80% of patients are included in these mutations. To make the reagents a kit, the procedure became simple and rapid. DNA was extracted by salting out method. The PCR product was detected by polyacrylamide gel electrophoresis and silver staining. The confirmation of the variant was done by the PCR-direct sequencing method. It took approximately six hours for PCR reaction, electrophoresis and staining. This method made us to detect and diagnose beta-Thal in one day.
authors.affiliation: Department of Clinical Nutrition, Faculty of Medical Health Science and Technology, Kawasaki University of Medical Welfare, Kurashiki 701-0193, Japan. haranok@bcc.kawasaki-m.ac.jp
authors.forename: Keiko
authors.initials: K
authors.lastname: Harano
authors.forename: Teruo
authors.initials: T
authors.lastname: Harano
mesh_terms: D005787:Gene Frequency / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D013997:Time Factors; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics