| 双通道荧光PCR快速检测水产品中主要致病性弧菌的研究 |
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| 引用本文： | 帅江冰,傅玲琳,张晓峰,励建荣.双通道荧光PCR快速检测水产品中主要致病性弧菌的研究[J].中国食品学报,2011,11(5). |
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| 作者姓名： | 帅江冰 傅玲琳 张晓峰 励建荣 |
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| 作者单位： | 1. 浙江出入境检验检疫局 杭州310012
2. 浙江工商大学食品与生物工程学院 浙江省食品安全重点实验室 杭州310035 |
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| 基金项目： | 浙江出入境检验检疫局科技项目(ZK200951); “十一五”国家863计划海洋技术领域重点项目(2007AA091806) |
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| 摘 要： | 目的:建立水产品中副溶血弧菌和霍乱弧菌快速、敏感、特异的双通道荧光PCR同步鉴别体系.方法:针对副溶血弧菌TDH基因和霍乱弧菌ctxA基因设计合成2对特异性引物和2条Taqman探针,优化体系,建立双通道荧光PCR体系.结果:建立的双通道荧光PCR体系特异性强,引物和探针之间无干扰.对副溶血孤菌和霍乱孤菌的检测灵敏度高,下限均能达到5× 104 CFU/mL.体系稳定,重复性好,可操作性强,两种不同浓度基因组DNA无相互抑制现象,适用于不同条件、多种样品的检测.采用该方法对140份采自舟山、象山和杭州的各类水产品进行检测,副溶血弧菌和霍乱孤菌检出率分别为25.7％和3.6％,未见霍乱弧菌和副溶血弧菌混合污染的样品,表明建立的双重荧光PCR是一种可用于水产品等样品中副溶血弧菌和霍乱弧菌的快速、灵敏、特异的鉴别方法.
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| 关 键 词： | 副溶血弧菌 霍乱孤菌 双通道荧光PCR Taqman探针 |
Study on the Duplex Real-time PCR Assay for Rapid Detection of Main Pathogenic Vibrio in Aquatic Products |
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| Shuai Jiangbing,Fu Linglin,Zhang Xiaofeng,Li Jianrong.Study on the Duplex Real-time PCR Assay for Rapid Detection of Main Pathogenic Vibrio in Aquatic Products[J].Journal of Chinese Institute of Food Science and Technology,2011,11(5). |
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| Authors: | Shuai Jiangbing Fu Linglin Zhang Xiaofeng Li Jianrong |
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| Affiliation: | Shuai Jiangbing1 Fu Linglin2* Zhang Xiaofeng1 Li Jianrong2(1Zhejiang Entry-Exit lnspection and Quarantine Bureau,Hangzhou 310012 2Food Safety Key Laboratory of Zhejiang Province,School of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310035) |
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| Abstract: | Objective:To develope a duplex real-time PCR assay for rapid detection of Vibrio cholerae and Vibrio parahaemolyticus simultaneously in aquatic products.Methods:two primer pairs and corresponding probes were designed against TDH gene of Vibrio parahaemolyticus and ctxA gene of Vibrio cholerae,respectively,to develop the duplex real-time assay.Results:the two pairs of primers and probes used in the assay were specific and had no cross-reations in detection of two genomic DNA.The duplex real-time PCR assay ha... |
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| Keywords: | Vibrio parahaemolyticus Vibrio cholerae duplex real-time PCR taqman probe |
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