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基于Cyt b的复杂生物样本中濒危动物的快速鉴别

Rapid Identification of Endangered Wildlife in Complex Biological Samples Based on Cyt b

  • 摘要: 本研究将二代高通量测序与DNA条形码技术相结合,通过人工制备多物种混合样本,以线粒体基因Cyt b为条形码,对混合样本中的动物种属进行同步鉴别,并确认濒危物种。结果表明,在属和种水平下,混合样本中物种的同步检出率均高达89%,而且灵敏度高,低至1%含量的物种成分被检出,但是种水平的物种分辨率并不高,近50%分类匹配为非目标匹配。本研究可得出,迷你型Cyt b片段可以作为混合生物样本中动物种属同步鉴别的条形码,在属水平下的鉴别率较高。在种水平下存在高检出、低分辨率的问题,后期可以通过增加条形码长度等方法进行进一步改善。本研究利用DNA宏条形码技术对条形码Cyt b在多物种混合生物样本中鉴别濒危动物的可行性进行评估,以期能为推进DNA宏条形码方法在动植物法医鉴定领域的常规化应用奠定前期基础。

     

    Abstract: This study combined the second generation sequencing with DNA barcoding technology. The mixed samples were manually prepared, and the mitochondrial gene Cyt b was used as a barcode to simultaneously identify the species in the mixed samples. The results showed that at the genus and species level, the identification rate of species in the mixed samples was 89%, and the sensitivity was high, as low as 1% of the species components were detected at the same time, but the species discrimination rate was low, nearly 50% of the taxonomy matches were non-target matches. The conclusion of this study was that the mini Cyt b fragment could be used as a barcode for simu-ltaneous identification of animal species in mixed biological samples. The discrimination rate was high at the genus level. At the species level, there was a high detection and low discrimination rate. This problem will be further improved by increasing the length of barcode. In this study, DNA metabarcoding technology was used to evaluate the feasibility of Cyt b in identifying endangered animals in multi-species mixed biological samples, in order to lay a preliminary foundation for the advancement of DNA metabarcoding method in the field of wildlife forensic identification.

     

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