National DNA Analysis Centre, Central Forensic Science Laboratory, 30-Gorachand Road, Kolkata-700 014, West Bengal, India.
Forensic Sci Int Genet. 2011 Jun;5(3):181-4. doi: 10.1016/j.fsigen.2010.03.001. Epub 2010 Apr 7.
Illegal trade of snake skin and uncontrolled hunting have instigated the extermination of many endangered snake species. Efforts to check illegal trade are often impeded due to lack of proper species identification methods. Hence, conservation strategies demand for authentic and quick identification techniques to trace the origin of the seized samples. This study employs DNA mini-barcoding as a method to identify some endangered snake species of India. We have designed two sets of novel primers for targeting regions within the mitochondrial Cytochrome Oxidase I gene to produce 175 bp and 245 bp amplicons. 175 bp fragment was amplified in all 11 snake species studied while the 245 bp amplicon was obtained in 10 species. DNA mini-barcodes recovered from these amplicons enabled the identification of snake species by retrieving the sequences available in public databases. The similarity scores ranging from 98 to 100% (98% taken as threshold value for species identification) signify the consistency of these mini-barcodes in snake species identification. Moreover, the results of the validation study confirm the effectiveness of the technique in forensic perspective, where the diagnostic morphological features of the seized sample are often missing.
非法的蛇皮贸易和无管制的捕猎行为已经促使许多濒危蛇类物种灭绝。由于缺乏适当的物种鉴定方法，打击非法贸易的努力常常受阻。因此，保护策略需要真实、快速的鉴定技术来追踪被查获样本的来源。本研究采用 DNA 微条形码技术来鉴定印度的一些濒危蛇类物种。我们设计了两套针对线粒体细胞色素氧化酶 I 基因内区域的新型引物，以产生 175bp 和 245bp 的扩增子。在研究的 11 种蛇中均扩增出 175bp 片段，而 245bp 扩增子则在 10 种蛇中获得。从这些扩增子中回收的 DNA 微条形码可通过检索公共数据库中可用的序列来识别蛇类物种。相似度得分在 98%至 100%之间（98%作为物种鉴定的阈值），表明这些微条形码在蛇类物种鉴定中的一致性。此外，验证研究的结果证实了该技术在法医学方面的有效性，因为被查获样本的诊断形态特征常常缺失。
