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利用基于纸张的微流控芯片(LOC)对三种 CITES 列出的鲨鱼进行遗传鉴定。

Genetic identification of three CITES-listed sharks using a paper-based Lab-on-a-Chip (LOC).

机构信息

Ecology & Environment Research Centre, Manchester Metropolitan University, Manchester, United Kingdom.

Colegio de Ciencias Biológicas y Ambientales, Universidad San Francisco de Quito (USFQ), Quito, Ecuador.

出版信息

PLoS One. 2024 Apr 4;19(4):e0300383. doi: 10.1371/journal.pone.0300383. eCollection 2024.

DOI:10.1371/journal.pone.0300383
PMID:38574082
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10994358/
Abstract

Threatened shark species are caught in large numbers by artisanal and commercial fisheries and traded globally. Monitoring both which shark species are caught and sold in fisheries, and the export of CITES-restricted products, are essential in reducing illegal fishing. Current methods for species identification rely on visual examination by experts or DNA barcoding techniques requiring specialist laboratory facilities and trained personnel. The need for specialist equipment and/or input from experts means many markets are currently not monitored. We have developed a paper-based Lab-on-a-Chip (LOC) to facilitate identification of three threatened and CITES-listed sharks, bigeye thresher (Alopias superciliosus), pelagic thresher (A. pelagicus) and shortfin mako shark (Isurus oxyrinchus) at market source. DNA was successfully extracted from shark meat and fin samples and combined with DNA amplification and visualisation using Loop Mediated Isothermal Amplification (LAMP) on the LOC. This resulted in the successful identification of the target species of sharks in under an hour, with a working positive and negative control. The LOC provided a simple "yes" or "no" result via a colour change from pink to yellow when one of the target species was present. The LOC serves as proof-of-concept (PoC) for field-based species identification as it does not require specialist facilities. It can be used by non-scientifically trained personnel, especially in areas where there are suspected high frequencies of mislabelling or for the identification of dried shark fins in seizures.

摘要

受威胁的鲨鱼物种被手工和商业渔业大量捕捞,并在全球范围内交易。监测渔业中捕捞和销售的鲨鱼物种以及《濒危野生动植物种国际贸易公约》(CITES)限制产品的出口,对于减少非法捕捞至关重要。目前的物种鉴定方法依赖于专家的目视检查或 DNA 条码技术,这些方法需要专门的实验室设施和训练有素的人员。由于需要专门的设备和/或专家的投入,许多市场目前都没有得到监测。我们开发了一种基于纸张的芯片实验室(LOC),以方便在市场来源处鉴定三种受威胁和 CITES 列出的鲨鱼,大眼长尾鲨(Alopias superciliosus)、远洋长尾鲨(A. pelagicus)和短鳍灰鲭鲨(Isurus oxyrinchus)。从鲨鱼肉和鳍样本中成功提取了 DNA,并与 LOC 上的环介导等温扩增(LAMP)结合进行 DNA 扩增和可视化。这导致在不到一个小时的时间内成功鉴定了目标鲨鱼物种,具有工作阳性和阴性对照。LOC 通过从粉红色变为黄色来提供目标物种存在的简单“是”或“否”结果。LOC 作为基于现场的物种鉴定的概念验证(PoC),因为它不需要专门的设施。它可以由非科学训练的人员使用,特别是在怀疑存在高频率标签错误的地区,或者用于鉴定缉获的干鲨鱼鳍。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1625/10994358/88e185448604/pone.0300383.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1625/10994358/7cdb2b2144de/pone.0300383.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1625/10994358/486b9c15f95c/pone.0300383.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1625/10994358/09fc4905a090/pone.0300383.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1625/10994358/88e185448604/pone.0300383.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1625/10994358/7cdb2b2144de/pone.0300383.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1625/10994358/486b9c15f95c/pone.0300383.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1625/10994358/09fc4905a090/pone.0300383.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1625/10994358/88e185448604/pone.0300383.g004.jpg

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