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首页> 外文期刊>FEMS immunology and medical microbiology >Application of real-time PCR for quantitative detection of Vibrio parahaemolyticus from seafood in eastern China.
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Application of real-time PCR for quantitative detection of Vibrio parahaemolyticus from seafood in eastern China.

机译：实时荧光定量PCR在中国东部海产品中副溶血性弧菌定量检测中的应用。

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摘要

Vibrio parahaemolyticus is recognized as a leading human food-borne pathogen. A TaqMan PCR assay based on the gyrase B gene (gyrB) sequence of V. parahaemolyticus was developed for quantitative detection of V. parahaemolyticus in seafood. The study involving 27 V. parahaemolyticus and 10 strains of other species indicated that the real-time PCR test was highly specific. The sensitivity of the assay was approximately a single CFU per PCR in pure culture and six to eight CFU per PCR in spiked raw oyster, respectively. Real-time PCR values of artificially inoculated oyster homogenates correlated well with plate counts determined using culture methods. A total of 300 seafood samples were analyzed and 78 (26%) of these samples were positive for V. parahaemolyticus using a conventional culture method and 97 (32.3%) using the real-time PCR assay. All culture-positive samples were PCR-positive. However, 19 samples positive by PCR were culture-negative. The results show that retail seafood is commonly contaminated with V. parahaemolyticus in harvest season in eastern China. These data also indicate that real-time PCR can provide sensitive species-specific detection and quantification of V. parahaemolyticus in seafood without prior isolation and characterization of the bacteria by traditional microbiological methods.
机译：副溶血性弧菌是公认的人类食源性致病菌。开发了一种基于副溶血弧菌的旋回酶B基因（gyrB）序列的TaqMan PCR检测方法，用于定量检测海鲜中的副溶血弧菌。涉及27株副溶血性弧菌和10株其他物种的研究表明，实时PCR测试具有很高的特异性。该测定法的灵敏度分别约为纯培养物中每个PCR单个CFU和加标生蚝中每个PCR六至八个CFU。人工接种的牡蛎匀浆的实时PCR值与使用培养方法确定的板数密切相关。总共分析了300个海鲜样品，使用常规培养方法对这些样品中的78个（26％）的副溶血弧菌呈阳性，使用实时PCR分析的样品中的97个（32.3％）。所有培养阳性样品均为PCR阳性。但是，PCR阳性的19个样品为培养阴性。结果表明，在中国东部的收获季节，零售海鲜通常被副溶血弧菌污染。这些数据还表明，实时PCR可以提供灵敏的物种特异性检测和定量海鲜中副溶血性弧菌，而无需通过传统的微生物学方法预先分离和鉴定细菌。

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