Tozzo Pamela, Ponzano Elena, Novelli Enrico, Onisto Maurizio, Caenazzo Luciana
Department of Environmental Medicine and Public Health, Legal Medicine Section, University of Padua, Padova, Italy.
Am J Forensic Med Pathol. 2011 Jun;32(2):180-2. doi: 10.1097/PAF.0b013e31820c2bba.
Identification of a report's species is one of the basic analyses in forensic laboratories. The authors report the case of 6 bone fragments recovered in a wooded area, which were not attributable to 1 animal species on the basis of morphologic examination. The aim of this study was to develop a duplex polymerase chain reaction (PCR) to discriminate human and animal origin of bone fragments. The method is based on the PCR amplification of cytochrome b and a 16S ribosomal mitochondrial DNA fragment, which has never been tested up to now. Our protocol combines a single-round PCR with direct visualization of amplicons in agarose gel, without sequencing analysis of the PCR products. The presence of a single band (359 bp) indicates a nonhuman origin of the sample, whereas 2 bands (157 and 359 bp) indicate a human biologic sample.This method revealed to be useful for forensic purposes because the 16S ribosomal mitochondrial DNA is a small human-specific fragment that is easily amplifiable even with degraded DNA from biologic materials such as old bones.
鉴定报告中的物种是法医实验室的基本分析之一。作者报告了在一片林区发现的6块骨头碎片的案例，根据形态学检查，这些碎片并非来自单一动物物种。本研究的目的是开发一种双重聚合酶链反应（PCR），以区分骨头碎片的人类和动物来源。该方法基于细胞色素b和一个16S核糖体线粒体DNA片段的PCR扩增，到目前为止从未进行过测试。我们的方案将单轮PCR与琼脂糖凝胶中扩增子的直接可视化相结合，无需对PCR产物进行测序分析。单一条带（359 bp）的出现表明样本来源非人类，而两条带（157和359 bp）则表明是人类生物样本。该方法被证明对法医鉴定有用，因为16S核糖体线粒体DNA是一个小的人类特异性片段，即使是来自陈旧骨头等生物材料的降解DNA也很容易扩增。
