| 牡蛎中副溶血弧菌荧光定量PCR检测方法的建立及其应用 |
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| 引用本文： | 林强,李宁求,付小哲,刘礼辉,石存斌,吴淑勤.牡蛎中副溶血弧菌荧光定量PCR检测方法的建立及其应用[J].中国水产科学,2011,18(1):96-102. |
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| 作者姓名： | 林强 李宁求 付小哲 刘礼辉 石存斌 吴淑勤 |
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| 作者单位： | 1. 中国水产科学研究院,珠江水产研究所,广东,广州,510380;上海海洋大学,水产与生命学院,上海,201306
2. 中国水产科学研究院,珠江水产研究所,广东,广州,510380 |
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| 基金项目： | 国家863计划项目，农业部公益性行业科研专项 |
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| 摘 要： | 以副溶血弧菌毒素调控基因(toxin regulations,toxR)作为靶标基因,设计特异性引物及TaqMan探针,以含toxR基因的质粒为模板,建立质粒拷贝数与CT值的标准曲线,分别采用含toxR基因质粒、纯培养的副溶血弧菌和添加副溶血弧菌的牡蛎(Ostrea)模拟样品进行灵敏度试验,结果表明,其灵敏度分别为15拷贝、18 CFU/mL和180CFU/mL.同一个样品的30次重复性试验表明,试验内及试验间的变异系数分别为0.95%和1.5%.结果显示,本研究建立的副溶血弧菌荧光定量PcR检测方法特异性强、灵敏度高、重复性好,可用于牡蛎等水产品中副溶血弧菌的定量检测.
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| 关 键 词： | 副溶血弧菌 荧光定量PCR toxR基因 |
| 修稿时间： | 2011/1/14 0:00:00 |
Development and application of a real-time PCR assay for detection of Vibrio parahaemolyticuss in oyster |
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| LIN Qiang,LI Ningqiu,FU Xiaozhe,LIU Lihui,SHI Cunbin,WU Shuqin.Development and application of a real-time PCR assay for detection of Vibrio parahaemolyticuss in oyster[J].Journal of Fishery Sciences of China,2011,18(1):96-102. |
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| Authors: | LIN Qiang LI Ningqiu FU Xiaozhe LIU Lihui SHI Cunbin WU Shuqin |
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| Institution: | LIN Qiang1,2,LI Ningqiu1,FU Xiaozhe1,LIU Lihui1,SHI Cunbin1,WU Shuqin1 1.Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510380,China,2.College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306 |
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| Abstract: | Vibrio parahaemolyticus(VP) is a pathogen that is the leading cause of shellfish-associated cases of bacterial food poisoning.To establish a real-time PCR for quantitative analysis of toxR gene,a pair of specific primers and TaqMan fluorescent probe of toxR gene of VP were designed.The positive recombinant plasmids of toxR gene served as templates to establish standard curve,which showed corresponding relationship between the copies of plasmids and threshold cycle(CT) values.The lowest limit detection was 1... |
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| Keywords: | Vibrio parahaemolyticus real-time PCR toxR gene |
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