甲基化特异性MLPA技术在Prader-Willi综合征诊断中的应用价值

Value of methylation-specific mutiplex ligation-dependent probe in the diagnosis of Prader-Willi syndrome

目的不同发病机制的Prader-Willi综合征(PWS)在临床表现、预后和遗传风险上均存在一定差异,目前临床常用的确诊方法甲基化特异性PCR(MS-PCR)不能区分发病机制,本研究采用甲基化特异性多重连接依赖的探针扩增(MS-MLPA)技术诊断PWS,探讨其在诊断以及分辨发病机制上的优势。方法采用系统对照的方法,取经临床MS-PCR检查的30例患儿的外周血样本,其中包括通过MS-PCR确诊为PWS的病例16例,阴性对照病例14例,重新提取DNA,采用MS-MLPA试剂盒Me028进行基因检测分析。结果 MS-MLPA检测结果与MS-PCR检测结果一致,且检测出16例PWS病例中4例源于母源性同源二倍体,12例源于父源性15q11-q13区域缺失。结论 MS-MLPA是能鉴别PWS发病机制的一种可靠的实验诊断方法。

Objective Prader-Willi syndrome（PWS） with different pathogenesis has different clinical manifestations,prognosis and genetic risks.Pathogenesis of the disease cannot be explained by conventional diagnostic method MS-PCR.This study employed methylation-specific multiplex ligation-dependent probe amplification（MS-MLPA） for the diagnosis of PWS in order to explore the role of this method in the diagnosis and assessment of pathogenesis of PWS. Methods A system antithetical method was employed.Peripheral blood samples were collected from 30 children for MS-PCR.Of the 30 children,16 were diagnosed with PWS by MS-PCR and the other 14 showed negative MS-PCR.MS-MLPA kit Me028 was used to detect DNA extracted from the 30 samples. Results The results showed by MS-MLPA and MS-PCR were identical.MS-MLPA demonstrated that 4 cases were maternal uniparental disomy and 12 cases were paternal dfeletion in 15q11-q13 region. Conclusions MS-MLPA is a reliable method of genetic testing for PWS which can distinguish pathogenesis of PWS.