MS-MLPA方法在Prader-Willi综合征及Angelman综合征基因诊断中的应用  

Methylation-specific multiplex Hgation-dependent probe amplification in diagnosis of Prader-Wiili syndrome and Angelman syndrome

作　　者：李美蓉[1] 王小竹[2] 刘晓燕[1] 杨艳玲[1] 包新华[1] 张月华[1] 熊晖[1] 钟南[2] 秦炯[1] 吴希如[1] 潘虹[1] 

机构地区：[1]北京大学第一医院儿科,100034 [2]北京大学医学部遗传中心

出　　处：《中华医学杂志》2008年第46期3257-3261,共5页National Medical Journal of China

摘　　要：目的检测甲基化特异性的多重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Angelman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ABI310进行基因型分析，最终所得数据用GeneMarkeT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。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 Peripheral 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 1 PWS patient）. DNA was extracted and purified. MS-MLPA was 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 cases with maternal 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 case 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.

关 键 词：Pmder-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 

分 类 号：R686[医药卫生—骨科学]