PDF(Pubmed)
Abstract：
Microhaplotypes (MHs) are widely accepted as powerful markers in forensic studies. They have the advantage of both short tandem repeats (STRs) and single nucleotide polymorphisms (SNPs), with no stutter and amplification bias, short fragments and amplicons, low mutation and recombination rates, and high polymorphisms. In this study, we constructed a panel of 50 MHs that are distributed on 21 chromosomes and analyzed them using the Multiseq multiple polymerase chain reaction (multi-PCR) targeted capture sequencing protocol based on the massively parallel sequencing (MPS) platform. The sizes of markers and amplicons ranged between 11-81 bp and 123-198 bp, respectively. The sensitivity was 0.25 ng, and the calling results were consistent with Sanger sequencing and the Integrative Genomics Viewer (IGV). It showed measurable polymorphism among sequenced 137 Southwest Chinese Han individuals. No significant deviations in the Hardy-Weinberg equilibrium (HWE) and linkage disequilibrium (LD) were found at all MHs after Bonferroni correction. Furthermore, the specificity was 1:40 for simulated two-person mixtures, and the detection rates of highly degraded single samples and mixtures were 100% and 93-100%, respectively. Moreover, animal DNA testing was incomplete and low depth. Overall, our MPS-based 50-plex MH panel is a powerful forensic tool that provides a strong supplement and enhancement for some existing panels.
摘要：
微单倍型（MHs）在法医研究中被广泛接受为强大的标记。它们具有短串联重复序列（STR）和单核苷酸多态性（SNP）的优势，没有口吃和放大偏差，短片段和扩增子，低突变率和重组率，和高度多态性。在这项研究中,我们构建了一组分布在21条染色体上的50个MHs,并使用基于大规模平行测序(MPS)平台的Multiseq多重聚合酶链反应(multi-PCR)靶向捕获测序方案进行了分析.标记和扩增子的大小在11-81bp和123-198bp之间。分别。灵敏度为0.25ng，并且调用结果与Sanger测序和整合基因组学查看器（IGV）一致。在137个中国西南汉族个体中显示出可测量的多态性。Bonferroni校正后，在所有MHs中均未发现Hardy-Weinberg平衡（HWE）和连锁不平衡（LD）的显着偏差。此外，模拟两人混合物的特异性为1:40，高度降解的单一样品和混合物的检出率分别为100%和93-100%，分别。此外，动物DNA测试不完整且深度低。总的来说,我们基于MPS的50-plexMH面板是一个强大的法医工具，为一些现有的面板提供了强大的补充和增强。
