利用实时荧光PCR鉴定小体鲟物种的快速方法

A rapid quantitative real-time PCR method for identification of Acipenser ruthenus

从小体鲟线粒体基因筛选出位于COI（细胞色素氧化酶）基因中的一段保守序列，针对其设计引物，优化SYBRGreen实时荧光PCR反应体系，建立了一种鉴定小体鲟的实时荧光PCR定性检测方法．该方法检测小体鲟DNA灵敏度为0．04mg／L．通过对采集和市售小体鲟鱼样品检测，该方法可检测出样品中的小体鲟成分．实验证明该方法可对血液样品和组织样品中的小体鲟进行种源鉴别．

A set of primers were designed according to the conserved sequence of gene CO l （cyto- chrome oxidase）in mitochondria of Acipenser ruthenus cells, and a RT-PCR method was established for quantitative detection of Acipenser ruthenus-derived components. The primers were highly specific for Acipenser ruthenus DNA. The limit of detection was 0.04 ng per reaction. Based on the assay of the commercial and collecting ruthenus samples,the method is proven to be valid for the detection of Acipenser ruthenus sturgeon. In summary, this RT-PCR proved to be a reliable approach in identifica tion of Acipenser ruthenus ingredients.