Expansion of a SNaPshot assay to a 55‐SNP multiplex: Assay enhancements, validation, and power in forensic science
Expansion of a SNaPshot assay to a 55‐SNP multiplex: Assay enhancements, validation, and power in forensic science
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DOI：
10.1002/elps.201500353
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发表时间：
2016-05
期刊：
影响因子：
2.9
通讯作者：
Qian Wang;Li-hong Fu;Xiao-jing Zhang;Xinyu Dai;Mei Bai;Guangping Fu;B. Cong;Shujin Li
中科院分区：
文献类型：
--
作者：
Qian Wang;Li-hong Fu;Xiao-jing Zhang;Xinyu Dai;Mei Bai;Guangping Fu;B. Cong;Shujin Li
将先前开发的具有44个个体识别SNP的多重测定扩展到55重测定。在一个PCR反应中扩增了54个高信息SNP和一个釉原蛋白性别标记，然后使用CE用两个SNaPshot反应进行检测。改变了4个基因座的PCR引物、28个单碱基延伸引物和反应条件，以提高方法的耐用性。开发了等位基因调用的详细方法来指导电泳图的分析。分析了中国河北汉族人群的180个无关个体和100个父亲-孩子-母亲三人组。SNP位点未发现突变。组合平均匹配概率和累积排除概率分别为1.327 × 10−22和0.999932。对54个SNP和26个STR（包括在AmpFLSTR Identifiler和Investigator HDplex试剂盒中）的分析显示没有显著的连锁不平衡。我们的研究表明，扩展SNP多重检测是一种简单易行和有价值的方法，以补充STR分析。
A previously developed multiplex assay with 44 individual identification SNPs was expanded to a 55plex assay. Fifty‐four highly informative SNPs and an amelogenin sex marker were amplified in one PCR reaction and then detected with two SNaPshot reactions using CE. PCR primers for four loci, 28 single‐base extension primers, and the reaction conditions were altered to improve the robustness of the method. A detailed approach for allele calling was developed to guide analysis of the electropherogram. One hundred and eighty unrelated individuals and 100 father‐child‐mother trios of the Han population in Hebei, China were analyzed. No mutation was found in the SNP loci. The combined mean match probability and cumulative probability of exclusion were 1.327 × 10−22 and 0.999932, respectively. Analysis of the 54 SNPs and 26 STRs (included in the AmpFLSTR Identifiler and Investigator HDplex kits) showed no significant linkage disequilibriums. Our research shows that the expanded SNP multiplex assay is an easily performed and valuable method to supplement STR analysis.