Detection of short tandem repeat polymorphisms from human nails using direct polymerase chain reaction method
Detection of short tandem repeat polymorphisms from human nails using direct polymerase chain reaction method
复制标题
直接聚合酶链反应法检测人指甲短串联重复多态性
DOI：
10.1002/elps.201400061
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发表时间：
2014
期刊：
影响因子：
2.9
通讯作者：
Seisaku Uchigasaki
中科院分区：
文献类型：
--
作者：
Jian Tie;Seisaku Uchigasaki
人指甲是大规模灾害中亲子鉴定和个体识别的重要法医学材料。从硬组织中检测STR多态性通常需要DNA纯化，这在技术上是复杂且耗时的。在本研究中，我们尝试使用GenePrint® SilverSTR® III系统或AmpF STR® Identifiler® PCR扩增试剂盒提供的引物混合物，以及已知可有效扩增粗品样本的Tks Gflex DNA聚合酶，通过直接PCR扩增方法检测未经处理的人指甲样本的STR多态性。将宽约1.5mm、长约0.5mm的指甲碎片直接放入PCR管中，并测试各种PCR条件。PCR产物经变性丙烯酰胺凝胶电泳或CE分析。成功检测到多个STR多态性。该方法不仅可以从新鲜指甲中检测STR多态性，而且可以从室温下保存长达10年的旧指甲碎片中检测STR多态性，有望成为法医学和遗传学研究中的一种新的DNA分析方法。
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.