Salian S, Vahab S A, Shah H, Shukla A, Shenoy R, Kamath N, Shenoy J, Satyamoorthy K, Girisha K M
Genet Couns. 2016;27(4):449-460.
We set out to evaluate multiplex ligation dependent probe amplification (MLPA) as a tool for diagnosis and carrier detection in families with a dystrophinopathy. Fifty three Indian families with provisional diagnosis of Duchene muscular dystrophy or Becker muscular dystrophy were evaluated by MLPA and multiplex polymerase chain reaction (PCR). Sanger sequencing was used to analyze the entire gene in one patient. Mothers were tested for carrier status whenever possible. Molecular analysis of DMD gene by combining MLPA and multiplex PCR yielded a mutation detection rate of 62% (33/53). Deletions were detected in 27/53 (51%) cases, duplications in 5/53 (9%) cases, a small deletion one case and Sanger sequencing detected a nonsense mutation in one case. Mutation was not detected in 36% (19/53) cases. Fifty six percent of mothers (9/16) were found to be carriers. MLPA helped to refine the results of multiplex PCR testing in 22 patients (5 duplications, 16 deletions and one small deletion). We also describe a situation where a deletion of single exon on MLPA (but not detected by multiplex PCR) was actually due to a deletion of two nucleotides in the probe ligation site. MLPA appears to score over multiplex PCR in diagnosis and carrier detection, specifically by detecting deletions and duplications that are not detected by traditional multiplex PCR.
我们着手评估多重连接依赖探针扩增技术（MLPA）作为诊断和检测肌营养不良症家族中携带者的工具。通过MLPA和多重聚合酶链反应（PCR）对53个初步诊断为杜氏肌营养不良症或贝克肌营养不良症的印度家庭进行了评估。对一名患者的整个基因进行了桑格测序分析。尽可能对母亲进行携带者状态检测。联合使用MLPA和多重PCR对DMD基因进行分子分析，突变检出率为62%（33/53）。在27/53（51%）的病例中检测到缺失，5/53（9%）的病例中检测到重复，1例为小缺失，桑格测序在1例中检测到无义突变。36%（19/53）的病例未检测到突变。56%的母亲（9/16）被发现为携带者。MLPA有助于优化22例患者（5例重复、16例缺失和1例小缺失）的多重PCR检测结果。我们还描述了一种情况，即MLPA检测到单个外显子缺失（但多重PCR未检测到）实际上是由于探针连接位点的两个核苷酸缺失所致。MLPA在诊断和携带者检测方面似乎优于多重PCR，特别是在检测传统多重PCR未检测到的缺失和重复方面。
