Tie Jian, Uchigasaki Seisaku
Division of Legal Medicine, Department of Social Medicine, Nihon University School of Medicine, Tokyo, Japan.
Electrophoresis. 2014 Nov;35(21-22):3188-92. doi: 10.1002/elps.201400061. Epub 2014 Aug 14.
Human nail is an important forensic material for parental testing and individual identification in large-scale disasters. Detection of STR polymorphism from hard tissues generally requires DNA purification, which is technically complicated and time consuming. In the present study, we attempted to detect STR polymorphisms from untreated human nail samples by direct PCR amplification method using the primer mixture supplied with the GenePrint® SilverSTR® III System or the AmpFℓSTR® Identifiler® PCR Amplification Kit, and Tks Gflex DNA polymerase known to be effective for amplification from crude samples. A nail fragment measuring approximately 1.5 mm in breadth and 0.5 mm in length was placed directly into a PCR tube, and various PCR conditions were tested. The PCR products were analyzed by denaturing acrylamide gel electrophoresis or CE. Multiple STR polymorphisms were detected successfully. This method that detects STR polymorphisms not only from fresh human fingernails, but also from old nail fragments stored at room temperature for up to 10 years is expected to become a novel DNA analytical method in forensic medicine and genetic studies.
人类指甲是亲子鉴定和大规模灾难中个体识别的重要法医物证。从硬组织中检测STR多态性通常需要进行DNA纯化,这在技术上既复杂又耗时。在本研究中,我们尝试使用GenePrint® SilverSTR® III系统或AmpFℓSTR® Identifiler® PCR扩增试剂盒提供的引物混合物,以及已知对粗样品扩增有效的Tks Gflex DNA聚合酶,通过直接PCR扩增法从未经处理的人类指甲样本中检测STR多态性。将一个宽度约为1.5毫米、长度约为0.5毫米的指甲碎片直接放入PCR管中,并测试了各种PCR条件。PCR产物通过变性丙烯酰胺凝胶电泳或毛细管电泳进行分析。成功检测到多个STR多态性。这种不仅能从新鲜人类指甲,还能从在室温下保存长达10年的陈旧指甲碎片中检测STR多态性的方法,有望成为法医学和遗传学研究中的一种新型DNA分析方法。