Y-STR genetic screening by high-resolution melting analysis
Y-STR genetic screening by high-resolution melting analysis
复制标题
通过高分辨率熔解分析进行 Y-STR 基因筛查
DOI：
10.4238/gmr.15017266
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发表时间：
2016-01-01
影响因子：
0.4
通讯作者：
Li, W. H.
中科院分区：
文献类型：
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作者：
Deng, J. Q.;Liu, B. Q.;Li, W. H.
目前，广泛应用于法医实践的基于自动毛细管电泳的短串联重复序列（STR）基因分型方法在某些情况下费力、耗时、昂贵且具有技术挑战性。因此，需要新的基于分子的策略来最终确定法医相关的生物证据。在这里，我们使用高分辨率熔化分析（HRM）对y染色体STR基因分型进行法医遗传筛查。在10个Y-STR基因座中，初步研究了熔化剖面在稀释度、灵敏度、鉴别能力等因素上的再现性。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品中表现出更高的均匀性，并且能够在低至0.25 ng的浓度下成功检测DNA。对于混合样品，基于参考样品的熔点曲线曲线判别效率高。进行了三重y染色体STR HRM分析，为进一步的研究（如多重HRM分析）提供了基础。人力资源管理方法是一个一步的应用程序，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分析是一种有用的筛选工具，可用于法医实践。
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.