We aimed to develop a multiplex PCR method for the simultaneous detection of the three most consumed commercial loaches in East Asia: Misgurnus mizolepis, Misgurnus anguillicaudatus, and Paramisgurnus dabryanus. Phylogenetic analysis of genetic divergence and a phylogenetic tree based on the cytochrome oxidase I gene enabled successful genetic identification of the 3 species of loach. Multiplex species-specific PCR (MSS-PCR) primers designed based on single nucleotide polymorphisms were able to differentiate the species and generate PCR products among species that differed in size by at least 150 bp. The lengths of the amplified fragments were 818 bp for P. dabryanus, 669 bp for M. mizolepis, and 486 bp for M. anguillicaudatus, which could be clearly distinguished by gel electrophoresis. The three sets of PCR primers were mixed and applied to simultaneously detect the three loaches. The MSS-PCR markers designed in this study will be useful for the detection and identification of M. mizolepis, M. anguillicaudatus, and P. dabryanus.

我们旨在开发一种多重 PCR 方法，用于同时检测东亚消费量最大的三种商业泥鳅：Mizolepis、Misgurnus anguillicaudatus和Paramisgurnus dabryanus。遗传分化的系统发育分析和基于细胞色素氧化酶 I 基因的系统发育树能够成功地对 3 种泥鳅进行遗传鉴定。基于单核苷酸多态性设计的多重物种特异性 PCR (MSS-PCR) 引物能够区分物种并在大小差异至少 150 bp 的物种中产生 PCR 产物。P的扩增片段长度为 818 bp 。dabryanus , 669 bp 为M .mizolepis和M的 486 bp 。anguillicaudatus , 可通过凝胶电泳清楚地区分。三组PCR引物混合后应用于同时检测三条泥鳅。本研究设计的 MSS-PCR 标记物将有助于M的检测和鉴定。咪唑匹斯，M . anguillicaudatus和P . 达布里亚努斯