中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2008年
46期
3257-3261
,共5页
李美蓉%王小竹%刘晓燕%杨艳玲%包新华%张月华%熊晖%钟南%秦炯%吴希如%潘虹
李美蓉%王小竹%劉曉燕%楊豔玲%包新華%張月華%熊暉%鐘南%秦炯%吳希如%潘虹
리미용%왕소죽%류효연%양염령%포신화%장월화%웅휘%종남%진형%오희여%반홍
Prader-Willi综合征%Angelman综合征%甲基化特异性的多重连接依赖的探针扩增%甲基化特异性聚合酶链反应
Prader-Willi綜閤徵%Angelman綜閤徵%甲基化特異性的多重連接依賴的探針擴增%甲基化特異性聚閤酶鏈反應
Prader-Willi종합정%Angelman종합정%갑기화특이성적다중련접의뢰적탐침확증%갑기화특이성취합매련반응
Prader-willi syndrome%Angelman syndrome%Methylation-specific multiplex ligation-dependent probe amplification: Methylation specific polymerage chain reaction
目的 检测甲基化特异性的多莆连接依赖的探针扩增(MS-MLPA)方法的敏感性和可靠性,以寻求一种简单、重复性好、精确度高的用于Prader-Willi综合征(PWS)和Angeiman综合征(As)的基因诊断方法.方法 DNA提取试剂盒提取基因组DNA,然后,用DNA纯化试剂盒进行纯化.用MS-MLPA试剂盒Me028对临床诊断的2例PWS和4例AS患儿进行基因检测分析.扩增的PCR产物用测序仪ABI 310进行基因型分析,最终所得数据用GeneMarker软件进行分析.MS-MLPA的检测结果用甲基化特异性聚合酶链反应进行验证.结果 4例AS中3例源于母源染色体15q11～q13缺失,1例源于父源染色体15q11～q13单亲二倍体或印记中心缺陷;2例PWS中1例源于父源染色体15q11-q13缺失,1例尚不能明确原因.结论 MS-MLPA是一种简单、高效、准确、可靠的基因检测方法.
目的 檢測甲基化特異性的多莆連接依賴的探針擴增(MS-MLPA)方法的敏感性和可靠性,以尋求一種簡單、重複性好、精確度高的用于Prader-Willi綜閤徵(PWS)和Angeiman綜閤徵(As)的基因診斷方法.方法 DNA提取試劑盒提取基因組DNA,然後,用DNA純化試劑盒進行純化.用MS-MLPA試劑盒Me028對臨床診斷的2例PWS和4例AS患兒進行基因檢測分析.擴增的PCR產物用測序儀ABI 310進行基因型分析,最終所得數據用GeneMarker軟件進行分析.MS-MLPA的檢測結果用甲基化特異性聚閤酶鏈反應進行驗證.結果 4例AS中3例源于母源染色體15q11～q13缺失,1例源于父源染色體15q11～q13單親二倍體或印記中心缺陷;2例PWS中1例源于父源染色體15q11-q13缺失,1例尚不能明確原因.結論 MS-MLPA是一種簡單、高效、準確、可靠的基因檢測方法.
목적 검측갑기화특이성적다보련접의뢰적탐침확증(MS-MLPA)방법적민감성화가고성,이심구일충간단、중복성호、정학도고적용우Prader-Willi종합정(PWS)화Angeiman종합정(As)적기인진단방법.방법 DNA제취시제합제취기인조DNA,연후,용DNA순화시제합진행순화.용MS-MLPA시제합Me028대림상진단적2례PWS화4례AS환인진행기인검측분석.확증적PCR산물용측서의ABI 310진행기인형분석,최종소득수거용GeneMarker연건진행분석.MS-MLPA적검측결과용갑기화특이성취합매련반응진행험증.결과 4례AS중3례원우모원염색체15q11～q13결실,1례원우부원염색체15q11～q13단친이배체혹인기중심결함;2례PWS중1례원우부원염색체15q11-q13결실,1례상불능명학원인.결론 MS-MLPA시일충간단、고효、준학、가고적기인검측방법.
Objective To verify the sensitivity and reliability of methylation-specific multiplex ligation-dependent probe amplification(MS-MLPA)and to develop a simple,accurate,reliability method of genetic diagnosis for AS and PWS.Methods Pefipherai blood samples were collected from 4 suspected AS patients,2 suspected PWS patients,2 normal persons,and 2 molecular biologically proven positive controls (1 AS patient and l PWS patient).DNA was extracted and purified.MS-MLPA wag used to detect the methylation of the CpG dinucleotide and the copy number in the 15q.q13 region.The results of MS-MLPA were confirmed by MSP.Results Three cages with matemal deletion on 15q11-q13 region and one case with paternal uniparental disomy(UPD)or imprinting center defect in 15q11-q13 region were found in the 4 suspected AS patients.One PWS cage was found to be with paternal deletion in 15q11-q13 region and the other with paternal deletion in 15q11-q13 region or UPD or imprinting center defect in 15q11-q13 region.Conclusion MS-MLPA is a simple.rapid,accurate, and reliable method of genetic test.
