【摘要】：目的建立熒光實(shí)時定量PCR(qPCR)定量檢測溶藻弧菌的方法,檢測該方法的靈敏度并與傳統(tǒng)細(xì)菌培養(yǎng)比較其特異性吻合率。方法選擇溶藻弧菌的鞭毛基因fli C為目的基因設(shè)計探針引物,建立Taqman探針qPCR體系。采用雙盲法設(shè)計,分別用qPCR和Biolog Microstation System對溶藻弧菌和10株相關(guān)細(xì)菌進(jìn)行鑒定,以考核qPCR檢測體系的特異性。同時,通過梯度稀釋和繪制標(biāo)準(zhǔn)曲線,評價利用qPCR檢測溶藻弧菌的靈敏度。結(jié)果本試驗(yàn)建立的qPCR方法能特異、準(zhǔn)確、快速鑒定溶藻弧菌,敏感度達(dá)102CFU/ml。結(jié)論 qPCR方法能夠有效快速檢測溶藻弧菌。
[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.
【作者單位】： 解放軍第一七五醫(yī)院/廈門大學(xué)附屬東南醫(yī)院;
【基金】：南京軍區(qū)醫(yī)學(xué)科技創(chuàng)新項(xiàng)目(項(xiàng)目編號:MS093)
【分類號】：R440

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