Terminal restriction fragment length polymorphism profiling of bacterial flora derived from single human hair shafts can discriminate individuals
Terminal restriction fragment length polymorphism profiling of bacterial flora derived from single human hair shafts can discriminate individuals
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DOI：
10.1016/j.legalmed.2017.01.002
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发表时间：
2017-03-01
期刊：
影响因子：
1.5
通讯作者：
Sakai, Kenji
中科院分区：
文献类型：
--
作者：
Nishi, Eiji;Watanabe, Kota;Sakai, Kenji
人类毛发是在许多犯罪现场最常遇到的微量物证。然而，由于其形态学检验的准确性低、短串联重复序列（STR）分型的检测率低以及线粒体DNA分析中的异质性问题，它们尚未在实际的刑事调查中得到有效利用。在此，我们通过比较毛发上细菌群落的图谱来检验个体鉴别的可能性。我们对毛发样本中扩增的细菌16S核糖体RNA（rRNA）基因进行了末端限制性片段长度多态性（T - RFLP）分析。与现有的利用毛根的STR分型方法相比，这种利用毛干的方法能够检测到稳定的细菌DNA。我们成功地从所有接受测试的志愿者的单根毛干中获得了T - RFLP图谱。这些图谱对每个个体具有特异性，从个体自身获得的多个图谱比来自不同个体的图谱具有更高的相似性。当6个月后再次采集时，从大多数志愿者的样本中都能稳定地获得这些个体特异性图谱。将采集的毛发样本在30℃下储存对于获得可重复的T - RF图谱是有效的。当将实验室采集的不明毛发样本与预先构建的数据库进行比较时，22根毛发中的17根被归到一小群人当中，包括相应的个体。这些结果表明，对犯罪现场发现的毛干上的细菌群落进行T - RFLP分析可为缩小嫌疑人范围提供有用信息。（C）2017爱思唯尔有限公司。保留所有权利。
Human hairs are the trace evidence most commonly encountered at many crime scenes. However, they have not been effectively utilized for actual criminal investigations because of the low accuracy of their morphological inspection, low detection rate of short tandem repeat (STR) typing, and the problem of heteroplasmy in mitochondrial DNA analysis. Here, we examined the possibility of individual discrimination by comparing profiles of bacterial flora on hair. We carried out the profiling of terminal restriction fragment length polymorphisms (T-RFLP) of the amplified bacterial 16S ribosomal RNA (rRNA) gene from hair samples. Compared with existing SIR typing methods that use hair roots, this method using hair shafts allowed the detection of stable bacterial DNA. We successfully obtained the T-RFLP profile from single hair shafts of all volunteers tested. The profiles were specific to each individual, and multiple profiles obtained from the individual him/herself showed higher similarity than those from different individuals. These individual-specific profiles were stably obtained from samples from most volunteers, when collected again after 6 months. Storage of the collected hair samples at 30 degrees C was effective for obtaining reproducible T-RF profiles. When unidentified hair samples collected in the laboratory were compared with a pre-constructed database, 17 of 22 hairs were assigned to a small group of people, including the corresponding individuals. These results show that T-RFLP analysis of bacterial flora on a hair shaft found at a crime scene could provide useful information for narrowing down a suspect. (C) 2017 Elsevier B.V. All rights reserved.