两对交叉引物PCR检测非缺失型α地中海贫血点突变的建立及临床应用  

Establishment and clinical application of PCR-CTPP method for detection of non-deletion alpha thalassemia point mutations

作　　者：朱莉雅 卿吉琳[2] 曹津萌 魏燕 赵艺莲 叶超 周晓兴 谢梦如 陈治中[4] ZHU Liya;QING Jilin;CAO Jinmeng;WEI Yan;ZHAO Yilian;YE Chao;ZHOU Xiaoxing;XIE Mengru;CHEN Zhizhong(Guangxi University of Chinese Medicine,Nanning,530200,China;Center for Reproductive Medicine and Genetics,the People's Hospital of Guangxi Zhuang Autonomous Region&Guangxi Academy of Medical Sciences;Guilin Medical University;Joint Inspection Center of Precision Medicine,the People's Hospital of Guangxi Zhuang Autonomous Region&Guangxi Academy of Medical Sciences;Guangxi Medical University)

机构地区：[1]广西中医药大学,南宁530200 [2]广西壮族自治区人民医院·广西医学科学院生殖医学中心 [3]桂林医学院 [4]广西壮族自治区人民医院·广西医学科学院精准联合检验中心 [5]广西医科大学

出　　处：《临床血液学杂志》2024年第6期373-379,共7页Journal of Clinical Hematology

基　　金：广西重点研发计划项目(No:2023AB22089)。

摘　　要：目的建立一种基于两对交叉引物PCR技术(PCR with confronting two-pair primers,PCR-CTPP)同时快速检测非缺失型α地中海贫血WS、QS和CS三种突变类型的方法及其在临床中的应用。方法根据α珠蛋白基因中发生WS、QS和CS突变的区域基因序列,分别针对单个位点设计两对引物即通用外引物和特异性引物,经过聚合酶链反应、电泳鉴别基因型,同时检测非缺失型α地中海贫血。将该方法与PCR反向斑点杂交(PCR-RDB)技术比较,验证其检测准确性。结果PCR-CTPP方法可用于准确检测非缺失型α地中海贫血的WS、QS和CS突变类型,结果显示与常规非缺失型α地中海贫血PCR-RDB检测方法一致。结论PCR-CTPP技术检测非缺失型α地中海贫血WS、QS、CS三种突变位点,简单快速、经济省力,可用于非缺失型α地中海贫血的快速筛查和分子流行病学研究,适合在一般实验室推广应用。适用于非缺失型α地中海贫血的人群筛查及婚前或产前筛查,适合各个层次医院应用。Objective To establish a method for the simultaneous rapid detection of three mutation types,WS,QS,and CS,in non-deletion alpha thalassemia based on PCR with confronting two-pair primers(PCR-CTPP)and its application in clinical practice.Methods Based on the gene sequences of the region of theαpearl protein gene where WS,QS,and CS mutations occur,two pairs of primers,universal external primers and specific primers,were designed for a single locus,respectively,and the genotypes were identified by a polymerase chain reaction and electrophoresis to detect the non-deletion type ofαthalassemia at the same time.The accuracy of the method was verified by comparing it with PCR-RDB.Results The PCR-CTPP method can be used to accurately detect the WS,QS,and CS mutation types in non-deletionαthalassemia,and the results showed agreement with the conventional PCR-RDB assay for non-deletionαthalassemia.Conclusion PCR-CTPP is a simple,rapid,and cost-effective technique for the detection of WS,QS,and CS mutation sites in non-deficiencyαthalassemia,which can be used for rapid screening and molecular epidemiological studies of non-deficiencyαthalassemia,and it is suitable for general laboratory applications.It is suitable for population screening for non-deletion alpha-thalassemia and premarital or prenatal screening and is suitable for application in hospitals at all levels.

关 键 词：非缺失型α地中海贫血 PCR-CTPP法 点突变 

分 类 号：R446.11[医药卫生—诊断学]