efficient detection of mediterranean β-thalassemia mutations by multiplex single-nucleotide primer extension有效的检测多路单核苷酸引物延伸的地中海β-thalassemia突变.pdfVIP

Efficient Detection of Mediterranean b-Thalassemia Mutations by Multiplex Single-Nucleotide Primer Extension 1 1 1 1,2 Biljana Atanasovska , Georgi Bozhinovski , Dijana Plaseska-Karanfilska , Lyubomira Chakalova * 1 Research Center for Genetic Engineering and Biotechnology, Macedonian Academy of Sciences and Arts, Skopje, Former Yugoslav Republic of Macedonia, 2 Department of Molecular Biology of the Cell Cycle, Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia, Bulgaria Abstract b-Thalassemias and abnormal hemoglobin variants are among the most common hereditary abnormalities in humans. Molecular characterization of the causative genetic variants is an essential part of the diagnostic process. In geographic areas with high hemoglobinopathy prevalence, such as the Mediterranean region, a limited number of genetic variants are responsible for the majority of hemoglobinopathy cases. Developing reliable, rapid and cost-effective mutation-specific molecular diagnostic assays targeting particular populations greatly facilitates routine hemoglobinopathy investigations. We developed a one-tube single-nucleotide primer extension assay for the detection of eight common Mediterranean b- thalassemia mutations: Codon 5 (-CT); CCT(Pro)-.C–, Codon 6 (-A); GAG(Glu)-.G-G, Codon 8 (-AA); AAG(Lys)-.–G, IVS-I-1 (G-.A), IVS-I-6 (T-.C), IVS-I-110 (G-.A), Codon 39 (C-.T), and IVS-II-745 (C- .G), as well as the hemoglobin S variant beta 6(A3) Glu.Val. We validated the new assay using previously genotyped samples obtaining 100% agreement between independent genotyping methods. Our approach, applicable in a range of Mediterranean countries, o