荧光实时定量PCR检测溶藻弧菌方法的建立
发布时间：2019-06-21 20:40
【摘要】：目的建立荧光实时定量PCR(qPCR)定量检测溶藻弧菌的方法,检测该方法的灵敏度并与传统细菌培养比较其特异性吻合率。方法选择溶藻弧菌的鞭毛基因fli C为目的基因设计探针引物,建立Taqman探针qPCR体系。采用双盲法设计,分别用qPCR和Biolog Microstation System对溶藻弧菌和10株相关细菌进行鉴定,以考核qPCR检测体系的特异性。同时,通过梯度稀释和绘制标准曲线,评价利用qPCR检测溶藻弧菌的灵敏度。结果本试验建立的qPCR方法能特异、准确、快速鉴定溶藻弧菌,敏感度达102CFU/ml。结论 qPCR方法能够有效快速检测溶藻弧菌。
[Abstract]:Objective to establish a fluorescence real-time quantitative PCR (qPCR) method for the quantitative detection of Vibrio alginolyticus, to detect the sensitivity of this method and to compare its specific coincidence rate with that of traditional bacterial culture. Methods the flagellum gene fli C of Vibrio alginolyticus was selected as the target gene to design probe primers to establish Taqman probe qPCR system. Vibrio alginolyticus and 10 strains of related bacteria were identified by qPCR and Biolog Microstation System, respectively, in order to assess the specificity of qPCR detection system. At the same time, the sensitivity of using qPCR to detect Vibrio alginolyticus was evaluated by gradient dilution and drawing standard curve. Results the qPCR method established in this experiment was specific, accurate and rapid for the identification of Vibrio alginolyticus with a sensitivity of 102 CFU / ml. Conclusion qPCR method can effectively and quickly detect Vibrio alginolyticus.
【作者单位】： 解放军第一七五医院/厦门大学附属东南医院;
【基金】：南京军区医学科技创新项目(项目编号:MS093)
【分类号】：R440
本文编号：2504334
[Abstract]:Objective to establish a fluorescence real-time quantitative PCR (qPCR) method for the quantitative detection of Vibrio alginolyticus, to detect the sensitivity of this method and to compare its specific coincidence rate with that of traditional bacterial culture. Methods the flagellum gene fli C of Vibrio alginolyticus was selected as the target gene to design probe primers to establish Taqman probe qPCR system. Vibrio alginolyticus and 10 strains of related bacteria were identified by qPCR and Biolog Microstation System, respectively, in order to assess the specificity of qPCR detection system. At the same time, the sensitivity of using qPCR to detect Vibrio alginolyticus was evaluated by gradient dilution and drawing standard curve. Results the qPCR method established in this experiment was specific, accurate and rapid for the identification of Vibrio alginolyticus with a sensitivity of 102 CFU / ml. Conclusion qPCR method can effectively and quickly detect Vibrio alginolyticus.
【作者单位】： 解放军第一七五医院/厦门大学附属东南医院;
【基金】：南京军区医学科技创新项目(项目编号:MS093)
【分类号】：R440
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