苏门答腊虎（Panthera tigris sumatrae，Pocock 1929）保存毛发和皮肤样本法医学研究DNA提取方法的发展

Q3 Agricultural and Biological Sciences

HAYATI Journal of Biosciences Pub Date : 2023-05-31 DOI:10.4308/hjb.30.5.816-824

N. Andayani, A. E. Maryanto, Muhammad Naufal Nur

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引用次数: 0

摘要

偷猎和非法野生动物贸易对苏门答腊虎构成了严重威胁。对老虎身体部位的高需求导致非法市场上出现大量仿制品，使没收案件的形态鉴定变得复杂。鉴于国家保护法只对印度尼西亚本土物种进行了监管，准确的识别在法律正当程序中至关重要。使用分子方法进行鉴定可以克服这个问题。然而，大多数非法交易的老虎身体部位都被保存了很长一段时间，这降低了回收DNA的数量和质量。本研究旨在开发一种快速有效的方法，从保存的法医样本中回收DNA。这些方法已经用从国家研究与创新局（BRIN，前身为LIPI）获得的几份博物馆砷处理过的毛发和一块虎皮样本、从明古鲁省自然资源保护局（BKSDA）获得的虎毛以及从BKSDA亚齐没收的虎皮样本进行了测试。使用离子交换、盐析和基于蛋白酶的方法提取DNA。结果表明，基于蛋白酶的提取优于其他提取方法，从保存的样品中获得了适用于基于PCR的物种鉴定的Cyt b和ND2 mtDNA部分基因的DNA分离株。然而，仍需要进一步的工作来恢复足够的DNA产量用于性别鉴定。

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Development of DNA Extraction Method for Forensics Studies of Preserved Hair and Skin Samples from Sumatran Tiger (Panthera tigris sumatrae, Pocock 1929)

Poaching and illegal wildlife trade present severe threats to the Sumatran tiger. The high demand for tiger body parts leads to a high number of imitations in illegal markets, complicating the morphological identification of any confiscation cases. Accurate identification is essential in legal due process, given that the national protection law only regulates Indonesia's native species. Identification using molecular approaches may overcome the problem. However, most illegally traded tiger body parts have been preserved for an extended period of time, reducing the quantity and quality of the recovered DNA. This study aimed to develop a fast and effective method to recover DNA from preserved forensic samples. The methods had been tested with several museum samples of arsenic-treated hairs and a tiger skin piece obtained from the National Research and Innovation Agency (BRIN, formerly LIPI), tiger hairs obtained from Conservation of Natural Resources Agency (BKSDA) of Bengkulu Province, and a confiscated tiger skin sample from BKSDA Aceh. The DNA was extracted using ion-exchange, salting out, and protease-based methods. The results showed that the protease-based extraction outperformed the others to yield applicable DNA isolates for PCR-based species identification by Cyt b and ND2 mtDNA partial genes from preserved samples. However, further works are still needed to recover sufficient DNA yields for sex identification.

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来源期刊

HAYATI Journal of Biosciences Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)

CiteScore

1.10

自引率

0.00%

发文量

审稿时长

24 weeks

期刊介绍： HAYATI Journal of Biosciences (HAYATI J Biosci) is an international peer-reviewed and open access journal that publishes significant and important research from all area of biosciences fields such as biodiversity, biosystematics, ecology, physiology, behavior, genetics and biotechnology. All life forms, ranging from microbes, fungi, plants, animals, and human, including virus, are covered by HAYATI J Biosci. HAYATI J Biosci published by Department of Biology, Bogor Agricultural University, Indonesia and the Indonesian Society for Biology. We accept submission from all over the world. Our Editorial Board members are prominent and active international researchers in biosciences fields who ensure efficient, fair, and constructive peer-review process. All accepted articles will be published on payment of an article-processing charge, and will be freely available to all readers with worldwide visibility and coverage.