4个Y-miniSTR基因座荧光标记复合扩增体系的建立

摘要

Objective:A quadruplex of primers aimed at 4 Y-chromosomal short tandem repeat ( Y-STR) loci ( DYS456, DYS392,DYS439,Y-GATA-H4) was built. The length of 4 amplicons was all shorter than 150bp which was called miniSTR. An exami-nation was carried out for sensitivity,DNA database compatibility and degraded DNA typing ability of the quardplex. Methods:4 prim-ers of Y-STR(DYS456,DYS392,DYS439,Y-GATA-H4) were specially selected for short amplicons. The front primer of DYS456, DYS392,DYS439,Y-GATA-H4 was labeled with FAM,JOE,ROX,TAMRA separately. DNA samples were collected from forensic ca-ses. And degraded samples were made by ultrasonic impact treatment. Ladder of the quadruplex was amplified from dilution of Y23 Lad-der with the quadruplex primers. The quadruplex was compared to promega Y23 for sensitivity,DNA database compatibility and highly degraded DNA typing ability. Results:Sensitivity of the quardplex was the same as Y23 in both common samples and highly degraded samples. As a result of test on degraded DNA samples using the quadruplex systems,the systems proved to be an more effective tools for analyzing degraded DNAs than Y23. And there were no differences in DNA typing results which means great DNA database compatibili-ty. Conclusion:The quadruplex of Y-miniSTR in addition to the commercial available Y-STR multiplex kits are highly useful for foren-sic practices of degraded DNA samples.%目的：：构建DYS456,DYS392,DYS439,Y-GATA-H4等4个Y-miniSTR基因座复合扩增体系,评价其检测敏感度、数据库兼容性和对降解检材的分型能力。方法：设计4个Y-miniSTR的引物,用FAM、JOE、ROX、TAMRA分别标记其上游引物,构建复合扩增体系。利用本体系的引物扩增稀释过的Y23试剂盒的ladder,制备等位基因分型标准物。比较该体系与Promega PowerPlex ® Y23试剂盒在普通检材、降解检材的分型能力、敏感度和分型结果。结果：同一样本体系分型结果和商业化试剂盒Y23 STR分型结果相同,4个Y-miniSTR复合扩增体系和对降解检材的分型成功率更高,敏感度一致,具有数据库兼容性。结论：本研究中的4个Y-miniSTR复合扩增体系对降解检材的检测具有应用价值,敏感性较好,具有数据库兼容性,可以作为对降解检材分型时商业化试剂盒的补充。