协和医学杂志Issue(4)：369-375,7.DOI:10.3969/j.issn.1674-9081.2014.04.003
甲基化特异性多重连接探针扩增及甲基化特异性PCR诊断Prader-Willi综合征方法比较
Comparison between Methylation-specific Multiplex Ligation-dependent Probe Amplification and Methylation-specific PCR in Diagnosis of Prader-Willi Syndrome
摘要
Abstract
Objective To compare the diagnostic effectiveness of methylation-specific PCR ( MS-PCR ) and methylation-specific multiplex ligation-dependent probe amplification ( MS-MLPA ) in clinical suspected Prader-Willi syndrome (PWS).Methods One hundred and two participants (57 males and 45 females) who visited Peking Union Medical College Hospital in the period from October 2005 to February 2014 were included in this retrospective study .Among the 102 participants , 16 were normal controls , 2 PWS cases confirmed by fluores-cence in situ hybridization or high resolutin banding were positive controls , and 84 were suspected PWS cases . Genomic DNA was extracted for MS-PCR and MS-MLPA, based on which diagnoses were made and genetic types distinguished .Chi-squared test was used to compare the sensitivity , specificity , and accuracy of the two meth-ods .Results MS-PCR results showed that all normal controls and positive controls were conform to their pheno -types .Thirty-nine in the 84 suspected patients were shown to be PWS , and the other 45 to be normal .MS-MLPA showed that in the 86 non-normal participants ( positive control and suspected cases ) , 29 had paternal deletions , 9 had maternal uniparental disomy ( mUPD) , 47 were normal;and the assay failed to produce effective result in 1 case due to the overlong DNA storage time .Two participants were diagnosed as PWS cases by MS-PCR but nor-mal by MS-MLPA.The diagnosis of PWS was ruled out by repeat MS-PCR after doubling DNA dosage .Combined with clinical manifestations , 39 participants were definitively diagnosed as PWS , the other 63 were not PWS .The false-positive rate of MS-PCR was 3.17% (2/63).The sensitivity, specificity, and accuracy of MS-PCR were 100%, 96.83%, and 98.03%, respectively; while those indicators of MS-MLPA were 97.43%, 100%, and 99.02%, respectively , showing no significant difference ( all P>0.05 ) .Conclusions Both MS-PCR and MS-MLPA have high sensitivity , specificity , and accuracy , thus are effective for PWS diagnosis .MS-MLPA can dis-tinguish paternal deletion and mUPD .Enough dose of DNA is requisite in MS-PCR to avoid false-positive results . Freshness of DNA samples is essential in MS-MLPA, which also needs more strict performance standards .We suggest a combination of MS-PCR and MS-MLPA in diagnosing PWS to ensure the accuracy of the result .关键词
Prader-Will综合征/甲基化特异性PCR/甲基化特异性多重连接探针扩增Key words
Prader-Will syndrome/methylation-specific PCR/methylation-specific multiplex ligation-dependent probe ampli-fication分类
医药卫生引用本文复制引用
王薇,魏珉,宋红梅,邱正庆,施惠萍,赵时敏..甲基化特异性多重连接探针扩增及甲基化特异性PCR诊断Prader-Willi综合征方法比较[J].协和医学杂志,2014,(4):369-375,7.基金项目
杨森科学研究委员会中国分会研究基金（2011-2012年）和北京协和医院中青年基金 （）
