摘要
目的:探讨突变扩增系统实时荧光定量扩增法(ARMS-qPCR)用于检测GJB2基因235delC突变引起的非综合征型耳聋。方法:以GJB2 235delC基因序列为靶基因,基于该位点的序列设计特异性引物,并运用ARMS-qPCR技术对GJB2 235delC基因位点进行检测,从而区分野生型、突变纯合型和突变杂合型。同时以GJB2 235delC型样本53例及耳聋基因室间质评标准质控物45例作对比验证。结果:ARMS-qPCR的方法能有效区分GJB2 235delC野生型、突变纯合型和突变杂合型,且具有良好的特异性。结论:本实验建立的ARMS-qPCR方法检测GJB2 235delC基因突变引起的非综合征型耳聋,简便快速,成本低,灵敏度高,特异性强。
Objective:To establish an amplification refractory mutation system-quantitative real-time PCR(ARMSqPCR)method for detecting the non-syndromic hearing loss associated with the GJB2 235delC gene mutation.Methods:The specific primer pairs targeting both wild-type(WT)and mutant alleles of the GJB2 gene were used to develop an ARMS-qPCR-based assay that distinguishes between the GJB2 wild type,235delC mutant homozygous type and mutant heterozygous type.At the same time,53 cases of GJB2 235delC type samples and 45 cases of deafness genes control were compared and verified.Results:The ARMS-qPCR method exhibited had the robust capability in differentiating the wild type,mutant homozygous,and mutant heterozygous variants of GJB2 235delC,and with the excellent specificity.Conclusion:The ARMS-qPCR method established for detecting non-syndromic deafness caused by the GJB2 235delC gene mutation in this study is simple,rapid,cost-effective,and high sensitivity and strong specificity.
作者
戴翔
李隽
蔡文倩
熊乾
DAI Xiang;LI Jun;CAI Wenqian;XIONG Qian(Wuhan Children's Hospital,Wuhan Maternaland Child Health Care Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei Province,430016)
出处
《中国计划生育学杂志》
2025年第6期1369-1374,共6页
Chinese Journal of Family Planning
基金
湖北省自然科学基金面上项目(2014CKB511)
武汉市卫计委科研项目(WX15C20)联合资助。
