Department of Pathophysiology and High Altitude Physiology, Third Military Medical University, Chongqing, PR China.
Clin Chem Lab Med. 2010 Apr;48(4):475-83. doi: 10.1515/CCLM.2010.097.
The identification of human mitochondrial DNA (mtDNA) sequence variations, especially single nucleotide polymorphisms (SNPs), is important for many applications. The PCR-ligase detection reaction (LDR) method can reduce false-positives and eliminate the need for both post-PCR and post-ligation purifications in SNP analyses. In addition, it has been successfully employed to detect point mutations in various nuclear genes. In this study, we used the PCR-LDR platform to characterize mtDNA SNPs.
Multiplex PCR-LDRs were used to genotype 19 mtDNA single nucleotide polymorphic sites from 812 samples. Performance of the method was assessed by direct sequencing of 44 samples.
We established an overall 97.4% success rate with 99.2% accuracy using the multiplex PCR-LDR methodology.
The PCR-LDR mtDNA genotyping technique is simple, highly accurate, has high-throughput, and is cost-effective. Therefore, this method is applicable to mtDNA haplotyping in various applications.
鉴定人类线粒体 DNA（mtDNA）序列变异，尤其是单核苷酸多态性（SNPs），对于许多应用非常重要。聚合酶链反应-连接酶检测反应（LDR）方法可以减少假阳性，并消除 SNP 分析中聚合酶链反应后和连接后纯化的需要。此外，它已成功用于检测各种核基因中的点突变。在本研究中，我们使用 PCR-LDR 平台来描述 mtDNA SNPs。
使用多重 PCR-LDR 对 812 个样本中的 19 个 mtDNA 单核苷酸多态性位点进行基因分型。通过对 44 个样本进行直接测序来评估该方法的性能。
我们使用多重 PCR-LDR 方法建立了总体成功率为 97.4%，准确率为 99.2%。
PCR-LDR mtDNA 基因分型技术简单、高度准确、高通量且具有成本效益。因此，该方法适用于各种应用中的 mtDNA 单体型分析。
