用于法医学的高变线粒体DNA区域的高分辨率熔解分析（HRM）
High-Resolution Melting (HRM) of Hypervariable Mitochondrial DNA Regions for Forensic Science.
作者信息
Dos Santos Rocha Alípio, de Amorim Isis Salviano Soares, Simão Tatiana de Almeida, da Fonseca Adenilson de Souza, Garrido Rodrigo Grazinoli, Mencalha Andre Luiz
机构信息
Departamento de Biofísica e Biometria, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, RJ, 20551-030, Brazil.
Polícia Civil do Estado do Rio de Janeiro, Instituto de Pesquisas e Perícias em Genética Forense-IPPGF, Rio de Janeiro, RJ, 20230-240, Brazil.
出版信息
J Forensic Sci. 2018 Mar;63(2):536-540. doi: 10.1111/1556-4029.13552. Epub 2017 Aug 23.
Forensic strategies commonly are proceeding by analysis of short tandem repeats (STRs); however, new additional strategies have been proposed for forensic science. Thus, this article standardized the high-resolution melting (HRM) of DNA for forensic analyzes. For HRM, mitochondrial DNA (mtDNA) from eight individuals were extracted from mucosa swabs by DNAzol reagent, samples were amplified by PCR and submitted to HRM analysis to identify differences in hypervariable (HV) regions I and II. To confirm HRM, all PCR products were DNA sequencing. The data suggest that is possible discriminate DNA from different samples by HRM curves. Also, uncommon dual-dissociation was identified in a single PCR product, increasing HRM analyzes by evaluation of melting peaks. Thus, HRM is accurate and useful to screening small differences in HVI and HVII regions from mtDNA and increase the efficiency of laboratory routines based on forensic genetics.
法医鉴定策略通常通过分析短串联重复序列（STR）来进行；然而，已为法医学提出了新的额外策略。因此，本文对用于法医分析的DNA高分辨率熔解（HRM）进行了标准化。对于HRM，使用DNAzol试剂从八名个体的黏膜拭子中提取线粒体DNA（mtDNA），通过PCR扩增样本并进行HRM分析，以识别高变（HV）区域I和II中的差异。为了确认HRM，对所有PCR产物进行DNA测序。数据表明，通过HRM曲线可以区分不同样本的DNA。此外，在单个PCR产物中鉴定出罕见的双重解离，通过评估熔解峰增加了HRM分析。因此，HRM对于筛选mtDNA的HVI和HVII区域中的微小差异是准确且有用的，并提高了基于法医遗传学的实验室常规工作效率。
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