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首页> 外文期刊>Protein & Cell >Development of a real time PCR assay for rapid detection of Vibrio parahaemolyticus from seafood
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Development of a real time PCR assay for rapid detection of Vibrio parahaemolyticus from seafood

机译：开发用于快速检测海鲜中副溶血性弧菌的实时PCR检测方法

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A real time PCR assay for the detection of Vibrio parahaemolyticus in seafood samples was developed using a novel specific target and a competitive internal amplification control (IAC). The specificity of this assay was evaluated using 390 bacterial strains including V. parahaemolyticus , and other strains belonging to Vibrio and non- Vibrio species. The real time PCR assay unambiguously distinguished V. parahaemolyticus with a detection sensitivity of 4.8 fg per PCR with purified genomic DNA or 1 CFU per reaction by counting V. parahaemolyticu s colonies. The assays of avoiding interference demonstrated that, even in the presence of 2.1 μg genomic DNA or 107 CFU background bacteria, V. parahaemolyticu s could still be accurately detected. In addition, the IAC was used to indicate false-negative results, and lower than 94 copies of IAC per reaction had no influence on the detection limit. Ninety-six seafood samples were tested, of which 58 (60.4%) were positive, including 3 false negative results. Consequently, the real time PCR assay is effective for the rapid detection of V. parahaemotyticus contaminants in seafood.
机译：使用新型特异性靶标和竞争性内部扩增对照（IAC），开发了用于检测海鲜样品中副溶血性弧菌的实时PCR检测方法。使用包括副溶血弧菌和其他属于弧菌和非弧菌种的390种细菌菌株评估了该测定法的特异性。实时PCR分析可清楚地区分副溶血性弧菌，通过对纯净的溶血弧菌菌落计数，每个PCR用纯化的基因组DNA的检测灵敏度为4.8 fg，或每个反应的检测灵敏度为1 CFU。避免干扰的分析表明，即使在存在2.1μg基因组DNA或10 <7> CFU背景细菌的情况下，副溶血弧菌仍然可以准确检测到。此外，IAC用于指示假阴性结果，每个反应的IAC拷贝数低于94不会影响检测限。测试了96种海鲜样品，其中58例（60.4％）为阳性，包括3例假阴性结果。因此，实时PCR测定法对于快速检测海鲜中的副嗜血杆菌很有效。

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