三引物PCR技术在强直性肌营养不良1型患者基因检测及产前诊断中的应用  

Application of triplet-primer PCR technology for the genetic testing and prenatal diagnosis of patients with Myotonic dystrophy type 1

作　　者：陈晨[1] 赵学潮 孟静静[1] 孔祥东[1] Chen Chen;Zhao Xuechao;Meng Jingjing;Kong Xiangdong(Genetic and Prenatal Diagnosis Center,the First Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450052,China)

机构地区：[1]郑州大学第一附属医院遗传与产前诊断中心,郑州450052

出　　处：《中华医学遗传学杂志》2024年第10期1182-1186,共5页Chinese Journal of Medical Genetics

基　　金：国家重点研发计划(2018YFC1002206-2)。

摘　　要：目的探讨三引物PCR(TP-PCR)技术在强直性肌营养不良症1型(DM1)患者分子检测和产前诊断中的应用价值。方法选取2018年5月至2022年10月在郑州大学第一附属医院遗传与产前诊断中心接受基因检测的48个DM1家系共60人作为研究对象。应用TP-PCR结合毛细管电泳的方法对受试者进行DMPK基因CTG重复数的检测,并对4个确诊家系进行产前诊断。本研究通过郑州大学第一附属医院医学伦理委员会的审查(伦理号:KS-2018-KY-36)。结果共检出52例DM1患者,多以肌无力、肌萎缩和肌强直为首发症状,并伴有肌强直样放电;部分患者合并有其他系统的异常。患者的DMPK基因CTG异常扩增数>50次,正常等位基因CTG重复数范围为5~18次,检出率最高的等位基因重复次数为6次(30.77%,16/52)。接受产前诊断的4个DM1家系中,1例胎儿未见异常,3例胎儿检测到CTG异常扩增(>50次),并被诊断为DM1患者。结论TP-PCR可快速准确诊断DM1患者,但当CTG异常扩增数高于50次时,该方法无法准确检出重复次数。ObjectiveTo explore the application of triplet-primer PCR(TP-PCR)for the genetic testing and prenatal diagnosis in patients with Myotonic dystrophy type 1(DM1).MethodsA total of 60 individuals from 48 pedigrees undergoing genetic testing at the Genetic and Prenatal Diagnosis Center of the First Affiliated Hospital of Zhengzhou University from May 2018 to October 2022 were selected as the study subjects.TP-PCR combined with capillary electrophoresis was applied to determine the number of CTG repeats of the DMPK gene,and prenatal testing was provided to four DM1 pedigrees.This study was approved by Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University(Ethics No.KS-2018-KY-36).ResultsA total of 52 DM1 patients were detected,mostly with muscle weakness,muscular atrophy and myotonia as the initial symptoms,along with typical myotonic potentials.Some patients also had abnormalities of other systems.The number of abnormal CTG repeats of the DMPK gene was>50,whilst the number of CTG repeats on the normal allele had ranged from 5 to 18.The number of the most common normal CAG repeats was 6(30.77%,16/52).Among the four DM1 pedigrees undergoing prenatal diagnosis,one fetus was healthy,whilst three fetuses were found to have abnormal CTG repeats(>50 times)and diagnosed with DM1.ConclusionTP-PCR can diagnose DM1 patients with speed and accuracy.However,this method cannot accurately determine the number of CTG repeats when it exceeds 50.

关 键 词：强直性肌营养不良1型 三引物PCR CTG三核苷酸重复 

分 类 号：R440[医药卫生—诊断学] R714.5[医药卫生—临床医学] R746.2