Microhaplotype identified and performed in genetic investigation using PCR-SSCP
Microhaplotype identified and performed in genetic investigation using PCR-SSCP
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使用 PCR-SSCP 在遗传研究中识别和执行微单倍型
DOI：
10.1016/j.fsigen.2017.01.008
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发表时间：
2017
影响因子：
3.1
通讯作者：
Zhang Lin
中科院分区：
文献类型：
--
作者：
Chen Peng;Zhu Jing;Pu Yan;Jiang Youjing;Chen Dan;Wang Hui;Mao Jiong;Zhou Bin;Gao Linbo;Bai Peng;Liang Weibo;Zhang Lin
最近引入的微单倍型基因座的概念在法医学中引起了注意。以往的研究一般是通过获取大量无关个体的SNPs的基因数据，然后通过统计和计算技术对微单倍型进行阶段化，从而估计等位基因频率。单个新个体的确定阶段要求较大的个体集合以前已经进行过基因分型。仅由目标个体拥有的稀有微单倍型或以前没有研究过的特定群体的微单倍型不太可能利用SNP的数据集进行准确的阶段性研究。因此，需要一种能够直接确定单个个体精确的微单倍型信息的方法。在本研究中，我们介绍了基于大规模并行测序技术(MiSeq)的单链测序技术和基于PCR的单链构象多态(SSCP)技术，它们具有简单、准确和经济的特点。结果表明，微单倍型比按座位划分的SNPs包含更多的多态信息(平均杂合度为0.61比0.41)。比较中国五个民族人群的微单倍型等位基因频率，发现汉族和维吾尔族人群的微单倍型等位基因频率分布存在显著差异。进一步的Fst值配对分析和分子方差分析(AMOVA)表明，维吾尔族与其他群体之间存在显著的群体分化。
The recently introduced concept of microhaplotype loci has attracted attention in forensics. Previous studies estimated the allele frequencies generally through obtaining genotypic data on the individual SNPs from a larger set of unrelated individuals then phasing microhaplotypes by statistical and computational techniques. Determining phase for a single new individual requires the larger set of individuals to have been genotyped previously. Rare microhaplotypes possessed only by the target individual or microhaplotypes private to a specific population not previously studied are unlikely to be accurately phased using data sets of SNPs. Thus, there is a demand for an approach that could directly determine a gain single individual’s precise microhaplotype information. In the present study, we introduced potential approaches of single chain sequencing based Massively Parallel Sequencing Technology (MiSeq) and PCR based Single Strand Conformational Polymorphism (SSCP) technology which was simple, accurate, and cost-effective. The results indicated that microhaplotypes contain much more polymorphic information than divided SNPs per locus (average heterozygosity of microhaplotype 0.61 VS SNPs 0.41). When microhaplotype allele frequencies were compared among five Chinese ethnic populations, significantly different distributions were found between the Han and Uyghur populations. Further analysis of pairwise Fst values and analysis of molecular variance (AMOVA), showed significant population differentiation between the Uyghur and other populations.