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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。
Objective To establish a quantitative real-time quantitative PCR (qPCR) method for the detection of Vibrio alginolyticus, to test the sensitivity of this method and to compare its specificity with traditional bacterial culture. Methods The flagella gene fli C of Vibrio alginolyticus was selected as the target gene to design probe primers and the Taqman probe qPCR system was established. The double-blind design was used to identify V. alginolyticus and 10 strains of bacteria using qPCR and Biolog Microstation System respectively to test the specificity of qPCR detection system. In the meantime, the sensitivity of detecting V. alginolyticus using qPCR was evaluated by gradient dilution and plotting a standard curve. Results The qPCR method established in this study can specifically, accurately and rapidly identify V. alginolyticus with a sensitivity of 102 CFU / ml. Conclusion The qPCR method can detect Vibrio alginolyticus efficiently and rapidly.