Deng J Q, Liu B Q, Wang Y, Liu W, Cai J F, Long R, Li W H
Department of Forensic Medicine, Hainan Medical University, Haikou, China.
Center of Reproductive Medicine, Affiliated Hospital of Hainan Medical University, Haikou, China.
Genet Mol Res. 2016 Feb 5;15(1):gmr7266. doi: 10.4238/gmr.15017266.
Currently, the widely used automated capillary electrophoresis-based short tandem repeat (STR) genotyping method for genetic screening in forensic practice is laborious, time-consuming, expensive, and technically challenging in some cases. Thus, new molecular-based strategies for conclusively identifying forensically relevant biological evidence are required. Here, we used high-resolution melting analysis (HRM) for Y-chromosome STR genotyping for forensic genetic screening. The reproducibility of the melting profile over dilution, sensitivity, discrimination power, and other factors was preliminarily studied in 10 Y-STR loci. The results showed that HRM-based approaches revealed more genotypes (compared to capillary electrophoresis), showed higher uniformity in replicate tests and diluted samples, and enabled successful detection of DNA at concentrations as low as 0.25 ng. For mixed samples, the melting curve profiles discriminated between mixed samples based on reference samples with high efficiency. The triplex Y-chromosome STR HRM assay was performed and provided a foundation for further studies such as a multiplex HRM assay. The HRM approach is a one-step application and the entire procedure can be completed within 2 h at a low cost. In conclusion, our findings demonstrate that the HRM-based Y-STR assay is a useful screening tool that can be used in forensic practice.
目前，在法医实践中广泛用于遗传筛查的基于自动毛细管电泳的短串联重复序列（STR）基因分型方法费力、耗时、昂贵，并且在某些情况下技术上具有挑战性。因此，需要新的基于分子的策略来最终鉴定与法医相关的生物证据。在此，我们使用高分辨率熔解分析（HRM）进行Y染色体STR基因分型以用于法医遗传筛查。在10个Y-STR位点初步研究了熔解曲线在稀释、灵敏度、鉴别力和其他因素方面的重现性。结果表明，基于HRM的方法揭示了更多的基因型（与毛细管电泳相比），在重复测试和稀释样本中显示出更高的一致性，并且能够成功检测低至0.25 ng浓度的DNA。对于混合样本，熔解曲线图谱能够高效地区分基于参考样本的混合样本。进行了三重Y染色体STR HRM分析，并为进一步研究如多重HRM分析提供了依据。HRM方法是一步应用，整个过程可以在2小时内以低成本完成。总之，我们的研究结果表明，基于HRM的Y-STR分析是一种可用于法医实践的有用筛查工具。
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