Centre of Forensic Science, WestChem, Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow, G1 1XW, United Kingdom.
Forensic Sci Int Genet. 2012 Mar;6(2):250-7. doi: 10.1016/j.fsigen.2011.06.001. Epub 2011 Jul 1.
The tiger (Panthera tigris) is currently listed on Appendix I of the Convention on the International Trade in Endangered Species of Wild Fauna and Flora; this affords it the highest level of international protection. To aid in the investigation of alleged illegal trade in tiger body parts and derivatives, molecular approaches have been developed to identify biological material as being of tiger in origin. Some countries also require knowledge of the exact tiger subspecies present in order to prosecute anyone alleged to be trading in tiger products. In this study we aimed to develop and validate a reliable single assay to identify tiger species and subspecies simultaneously; this test is based on identification of single nucleotide polymorphisms (SNPs) within the tiger mitochondrial genome. The mitochondrial DNA sequence from four of the five extant putative tiger subspecies that currently exist in the wild were obtained and combined with DNA sequence data from 492 tiger and 349 other mammalian species available on GenBank. From the sequence data a total of 11 SNP loci were identified as suitable for further analyses. Five SNPs were species-specific for tiger and six amplify one of the tiger subspecies-specific SNPs, three of which were specific to P. t. sumatrae and the other three were specific to P. t. tigris. The multiplex assay was able to reliably identify 15 voucher tiger samples. The sensitivity of the test was 15,000 mitochondrial DNA copies (approximately 0.26 pg), indicating that it will work on trace amounts of tissue, bone or hair samples. This simple test will add to the DNA-based methods currently being used to identify the presence of tiger within mixed samples.
老虎（Panthera tigris）目前被列入《濒危野生动植物种国际贸易公约》附录 I，受到最高级别的国际保护。为了调查涉嫌非法交易老虎身体部位和衍生品的案件，分子方法已被开发用于鉴定生物材料是否来自老虎。一些国家还要求了解确切的老虎亚种，以便对涉嫌交易老虎产品的任何人进行起诉。在这项研究中，我们旨在开发和验证一种可靠的单一测定方法，以同时鉴定老虎物种和亚种；该测试基于鉴定老虎线粒体基因组中的单核苷酸多态性（SNP）。获得了目前野生存在的五个假定老虎亚种中的四个的线粒体 DNA 序列，并将其与来自 GenBank 的 492 只老虎和 349 种其他哺乳动物的 DNA 序列数据相结合。从序列数据中总共鉴定出 11 个 SNP 位点适合进一步分析。五个 SNP 是老虎特有的，六个 SNP 扩增了一个老虎亚种特有的 SNP，其中三个 SNP 是苏门答腊虎特有的，另外三个 SNP 是孟加拉虎特有的。多重测定能够可靠地鉴定 15 个凭证老虎样本。该测试的灵敏度为 15,000 个线粒体 DNA 拷贝（约 0.26 pg），表明它可以用于微量组织、骨骼或毛发样本。这种简单的测试将增加目前用于鉴定混合样本中老虎存在的基于 DNA 的方法。
