花胶鱼类物种Species-specific PCR和Multiplex PCR鉴定体系研究

China is the biggest global trader of fish maw (dried fish swimbladders). Over 100 fish maw species are commercially exploited all over the world, which export to China after capture and processed. It is very difficult to identify clearly the species of processed fish maws on the basis of their swimbladders morphology. Forensically Informative Nucleotide Sequencing (FINS) and DNA barcoding are effective for the detection of species adulteration from food products. However, it is difficult to implement these technologies for identification of mixed species. Species-specific PCR and Multiplex PCR have been proved to be effective alternatives in the identification of mixed species. Our previous studies showed that fish maw products were species substituted or unclear. Moreover, endangered species, protected species and toxic species were found in the market. The aim of this work is initially to establish the FINS and DNA barcoding method system for species identification of global fish maws and the DNA barcodes database, then carry out the research on evaluation of the labeling situation of fish maw products in the market, the global resources distribution mapping, global catch data statistics, endangered species, protected species and toxic species. On this base, Species-specific PCR and Multiplex PCR method system will be explored. The system is widely applicable, accurate, simple, fast, highly sensitive and easy to be standardized. The present study will provide the technical support and research approach for authentication and traceability of fish maw species, species diversity, resources analysis and protection of endangered species to conserve stocks biodiversity.

我国是全球花胶（鱼鳔干制品）第一贸易大国，全球花胶鱼类100余种，捕捞加工后输入我国，仅凭借鱼鳔形态特征很难辨认其真实物种。FINS和DNA条形码技术在物种鉴定方面展现巨大潜力，然而面对混合样品物种的鉴定难以实施。种特异性PCR和多重PCR技术给混合样品物种鉴定提供了新的思路。我们前期研究表明花胶存在物种替代和物种不明现象，更重要的，发现了濒危保护物种和毒性物种。本项目拟先进行全球花胶FINS、DNA条形码鉴定研究，建立DNA条形码数据库，评估我国市场花胶物种来源和水产品贴标现状，绘制全球自然资源分布图，统计全球捕捞量、揭示濒危保护物种和毒性物种。在此基础上，进行种特异性PCR和多重PCR鉴定体系探索研究，该体系具有适应范围广、准确、简便、快速、灵敏度高和易标准化的特点。本研究可为花胶物种多样性研究、物种溯源、种质资源分析和水产资源保护等方面提供技术支撑和研究途径。

我国是全球花胶（鱼鳔干制品）第一贸易大国，全球花胶鱼类100余种，捕捞加工后输入我国，仅凭借鱼鳔形态特征很难辨认其真实物种。FINS和DNA条形码技术在物种鉴定方面展现巨大潜力，然而面对混合样品物种的鉴定难以实施。种特异性 PCR和多重 PCR技术给混合样品物种鉴定提供了新的思路。本项目进行全球花胶FINS、DNA条形码鉴定研究，共采集获得花胶鱼类标准品103种、样品2336份，建立DNA条形码数据库，评估我国市场花胶物种来源和水产品贴标现状，绘制全球自然资源分布图，统计全球捕捞量、揭示濒危保护物种和毒性物种。在此基础上，进行种特异性 PCR和多重 PCR鉴定体系探索研究，设计了Species-specific PCR引物139对，设计了Multiplex PCR引物23组。该体系具有适应范围广、准确、简便、快速、灵敏度高和易标准化的特点。项目研究期间，共发表SCI论文7篇。本研究可为花胶物种多样性研究、物种溯源、种质资源分析、水产资源保护等方面提供技术支撑和研究途径。

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