Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw ul. Pawińskiego 5A, 02-106 Warsaw, Poland.
Gene. 2012 Sep 10;506(1):161-5. doi: 10.1016/j.gene.2012.06.081. Epub 2012 Jul 2.
Deletions in mitochondrial DNA are a common cause of mitochondrial disorders. The molecular diagnosis of mtDNA deletions for years was based on Southern hybridization later replaced by PCR methods such as PCR with primers specific for a particular deletion (mainly the so-called common deletion of 4977 bp) and long PCR. In order to evaluate the usefulness of MLPA (Multiplex Ligation-dependent Probe Amplification) in molecular diagnosis of large scale mtDNA deletions we compare four diagnostic methods: Southern hybridization, PCR, long-PCR and MLPA in a group of 16 patients with suspected deletions. Analysis was performed on blood, muscle and in one case hepatic tissue DNA. The MLPA was not able to confirm all the deletions detected by PCR methods, but due to its relative ease of processing, minimal equipment, low costs and the additional possibility to detect frequent point mtDNA mutations in one assay it is worth considering as a screening method. We recommend to always confirm MLPA results by PCR methods.
线粒体 DNA 缺失是线粒体疾病的常见原因。多年来，mtDNA 缺失的分子诊断基于 Southern 杂交，后来被 PCR 方法取代，如针对特定缺失的引物 PCR（主要是所谓的 4977bp 常见缺失）和长 PCR。为了评估 MLPA（多重连接依赖性探针扩增）在大规模 mtDNA 缺失分子诊断中的有用性，我们将 Southern 杂交、PCR、长 PCR 和 MLPA 这四种诊断方法在一组 16 名疑似缺失的患者中进行了比较。分析在血液、肌肉和一例肝组织 DNA 上进行。MLPA 无法确认所有通过 PCR 方法检测到的缺失，但由于其处理相对简单、设备要求低、成本低，并且可以在一次检测中检测到常见的点 mtDNA 突变，因此值得考虑作为一种筛选方法。我们建议始终通过 PCR 方法确认 MLPA 结果。
