Department of Molecular Pathology, Metropolis Healthcare Ltd., Mumbai, Maharashtra, India.
Department of Medical Genetics, Metropolis Healthcare Ltd., Mumbai, Maharashtra, India.
Neurol India. 2022 Jul-Aug;70(4):1615-1617. doi: 10.4103/0028-3886.355142.
Oftentimes, a variation at the multiplex ligation-dependent probe amplification (MLPA) probe binding site leads to improper hybridrization/ligation of the probe showing up as a deletion of an exon leading to false positive results for the detection of Duchenne muscular dystrophy (DMD)/Becker muscular dystrophy (BMD).
Investigating cases with single exon deletion using an alternate method [polymerase chain reaction (PCR) or sequencing] for confirmation of the deletion.
We evaluated males with single exon deletion (n = 49) in our study population (2015-2019). Forty-six were confirmed by an alternate method (conventional PCR/Sanger's sequencing) to confirm the deletion.
We observed 25.12% single exon deletions in our study cohort. Further evaluation determined a false positive rate of 6.12%. Three out of 49 single exon deletions had a point mutation near the probe-binding site, indicating a false positive result. Single exon deletions, thus, need to be evaluated with extreme caution, and point mutations, if any, need to be characterized to determine the nature of their pathogenicity.
在多重连接依赖性探针扩增（MLPA）探针结合位点中，经常会出现变异，导致探针杂交/连接不当，表现为外显子缺失，从而导致假阳性结果，从而导致假阳性结果检测杜氏肌营养不良症（DMD）/贝克肌营养不良症（BMD）。
使用替代方法（聚合酶链反应（PCR）或测序）对具有单外显子缺失的病例进行检测，以确认缺失。
我们评估了我们研究人群（2015-2019 年）中具有单外显子缺失的男性（n = 49）。46 例通过替代方法（常规 PCR/Sanger 测序）确认缺失。
我们观察到我们的研究队列中有 25.12%的单外显子缺失。进一步评估确定假阳性率为 6.12%。在 49 例单外显子缺失中有 3 例在探针结合位点附近存在点突变，表明结果为假阳性。因此，需要非常小心地评估单外显子缺失，并且如果有任何点突变，则需要对其进行特征描述，以确定其致病性。
