α地中海贫血的基因诊断技术研究

Study on microarray technique for rapid genetic diagnosis of α-thalassemia

目的建立一套设有内对照的、简便可靠的、诊断广东省常见的3种缺失型α地中海贫血的基因芯片技术,用于α地中海贫血的分子诊断。方法自行设计5组引物,优化PCR反应条件,运用PCR,芯片杂交,根据荧光信号的有无及位置检测中国人常见的3种缺失型(--SEA,-α3.7,-α4.2)α地中海贫血。结果成功地检测中国人常见的3种缺失型α地中海贫血,检测结果与Southern blotting分析及直接测序的结果一致。结论本研究所建立的方法准确、重复性好,便于临床样本分析及人群中α地中海贫血基因的筛查。

Objective：To develop a simple and reliable microarray technique for diagnosis of three commonest deletional（——SEA,-α4.2,-α3.7） and α-thalassemia gene in China.Methods：Five pairs of polymerase chain reaction（PCR） primers were designed by ourselves and used them to amplify the——SEA gene,-α4.2 gene and its normal control gene,-α3.7 gene and its normal control gene,and two respectively under an optimized PCR condition followed by microarray hybridization and fluorescent scanning.Finally,three commonest deletional α-thalassemia gene were identified according to the power of the fluorescence.Results：Heterozygotes,homozygotes and dual heterozygotes of the three deletional α-thalassemia mutations were successfully detected by the developed technique.The results were identical to those from Southern blotting analysis and DNA sequencing.Sixty-five cases of α-thalassemia were diagnosed by this technique.Conclution：The usage of microarray in identifying α-thalassemia mutations has the advantages of simplicity,reproducibility and high through-up.This technique does not use radioisotope and may be suitable for prenatal diagnosis and population screening.