预包装柳州螺蛳粉沙门氏菌实时荧光PCR快速检测

Rapid detection of Salmonella in prepackaged Liuzhou river snail rice noodles by real-time PCR

中文关键词:  预包装柳州螺蛳粉  沙门氏菌  实时荧光PCR

英文关键词:prepackaged Liuzhou river snail rice noodles  Salmonella  real-time PCR

中文摘要:

      目的 建立实时荧光PCR法快速检测预包装柳州螺蛳粉中沙门氏菌的分析方法。方法 根据沙门氏菌invA基因设计引物和探针, 优化反应体系中的探针浓度后对人工添加沙门氏菌和干扰菌模拟受污染的预包装柳州螺蛳粉样品进行检测。结果 设计的引物和探针只对阳性菌株有荧光反应, 具有良好的特异性, 最佳反应探针终浓度为0.2 μmol/L, 检测灵敏度为100 CFU/mL, 扩增效率103.92%; 对模拟污染的预包装柳州螺蛳粉经过一步增菌18 h后最低能检测出4 CFU/25 g沙门氏菌。结论 实时荧光PCR检测灵敏度高、特异性强, 可应用于预包装柳州螺蛳粉中沙门氏菌的快速高效检测。

英文摘要:

      Objective To establish a method for rapid detection of Salmonella in pre-packaged liuzhou snail meal by real-time PCR. Methods Primers and probes were designed according to the invA gene of Salmonella, and the concentration of the probes in the reaction system was optimized to detect the artificially added samples of contaminated pre-packaged Liuzhou snail meal with Salmonella and interfering bacteria. Results The primers and probes designed only had fluorescence response to the positive strain and had good specificity, the final concentration of the probe for the optimal reaction was 0.2 μmol/L and the achieved 100 CFU/mL detection sensitively and 103.92% amplification efficiently. The detection limit of the samples previously mentioned was 4 CFU/25 g (Salmonella) after one-shot bacterial proliferation with 18 h. Conclusion The real-time PCR with high sensitivity and specificity can be widely applied to detect the Salmonella in prepackaged Liuzhou river snail rice noodles effectively and rapidly.