高分辨率熔解曲线分析技术及其在法医学中的应用

High-resolution melting analysis in forensic research

高分辨率熔解曲线(high-resolution melting,HRM)分析技术是一种利用DNA杂交与退火熔解特性进行核酸分析的技术,主要通过对DNA解离活动中荧光染料变化的监控,实现突变的筛查和检测。HRM分析技术不需要探针标记,并且能够在很大程度上减少测序的负担,具有特异性好、通量高、时间成本低、便于操作等优点。HRM分析技术还是一种同质闭管的检测方法,聚合酶链式反应(polymerase chain reaction,PCR)与后续检测均在同一孔内完成,无需转移样品,能有效的减少交叉污染的可能性。近年来有越来越多的专家学者尝试将HRM分析技术应用于法医学研究和相关案例中。本文拟对高分辨率熔解曲线分析技术的原理、技术特点、局限性以及在法医学中的应用进行综述。

High-resolution melting （HRM） analysis is a versatile method for variant scanning and genotyping. It involves amplification of the target in the presence of a saturation dye by the polymerase chain reaction （PCR）. HRM analysis can be performed in one closed-tube, which does not require additional post-PCR separations and greatly reduces the possibility of contamination. HRM is faster, simpler, and less expensive than alternative approaches requiring labeled probes. Considering the many advantages of HRM analysis, many researchers have tried to apply this method to forensic research. This paper intends to summarize the principle, technical characteristics, limitations and application of HRM analysis in forensic science.