荧光定量聚合酶链式反应种植前胚胎β-地中海贫血基因诊断  

Preimplantation genetic diagnosis of β-thalassemia by real-time quantitative PCR

作　　者：李晓红[1] 庄广伦[2] 周灿权[2] 程钢[2] 舒益民[2] 曾瑞萍[2] 

机构地区：[1]北京大学第一医院生殖与遗传医疗中心,北京100034 [2]中山医科大学第一附属医院生殖中心

出　　处：《中国生育健康杂志》2003年第4期235-237,共3页Chinese Journal of Reproductive Health

基　　金：国家"973"重大基金资助项目 (批准号 :0 0 1cb5 10 10 2 0 1) ;国家自然科学基金资助项目 (批准号 :3 0 0 0 0 875及3 0 170 3 94);北京大学"985"基金资助项目

摘　　要：目的　用荧光定量聚合酶链式反应 (PCR)方法进行种植前β-地中海贫血一种中国人常见突变类型 CD4 1/ 4 2 (- TCTT)的基因诊断。　方法　来源于β-地中海贫血病人或携带者的 2 89个淋巴细胞和人体外受精 -胚胎移植志愿者剩余的胚胎分离 137个单细胞用荧光定量 PCR方法进行分析。　结果　用单个淋巴细胞诊断β-地中海贫血扩增效率达 96 % ,准确率达 99%～ 10 0 % ,等位基因丢失 (ADO)率仅 0～ 5 %。单个胚胎细胞扩增效率 86 %。　结论　荧光定量 PCR技术是一种敏感、快速、可靠和准确的技术 ,适合包括β-地中海贫血在内的单基因疾病的种植前诊断。Objective A recently developed system for fluorescence real-time quantitative polymerase chain reaction(PCR) to detect three types of β-thalassemia mutations[CD41/42 (-TCTT)], which accounted for about 40 % of all β-thalassemia alleles in Chinese and to prepare for clinical PGD of β-thalassemia. Methods 289 lymphocytes of β-thalassemia patients or carriers and 137 human blastomeres were analyzed by real-time quantitative PCR. Results Real-time quantitative PCR has high sensitivity,high reliability (96 %),and high accuracy(99 %～100 %)rates for β-thalassemia diagnosis in single lymphocytes and can reduce allelic dropout (ADO) rates to 0～5 %.High reliability (86 %) have been achieved in human blastomeres.All the results were confirmed by DNA amplification and reverse dot blot hybridization. Conclusion Real-time quantitative PCR proves to be a sensitive,convinient,reliable and accurate technique for PGD of β-thalassemia as well as other monogenic disorders.

关 键 词：荧光定量聚合酶链式反应 种植 胚胎 Β-地中海贫血 基因诊断 

分 类 号：R446.9[医药卫生—诊断学]