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目的通过建立霍乱弧菌的快速检测方法,提高霍乱疫情应急检测的速度和检出率,溯源可疑的传染源。方法将细菌分离鉴定,实时荧光PCR快速检测,胶体金免疫层析快速试纸法同时用于检测霍乱疫情处置期间所采集的患者水样便、外环境水质、食品及水产品,并对其结果进行比较分析;将分离到的O139霍乱弧菌进行脉冲场凝胶(PFGE)电泳分型分析。结果 3种检测方法中实时荧光PCR快速检测和胶体金免疫层析快速试纸法阳性率较高,常规细菌分离鉴定阳性率较低,3种方法各有优势及缺点;PFGE分子分型分析提示本次O139霍乱疫情传染源为水产市场中甲鱼带菌,污染病人食用食物所致。结论荧光PCR快速检测和胶体金免疫层析快速试纸法是菌株分离培养方法的有效补充;PFGE对不同来源的霍乱弧菌遗传同源相关性分析有较好的分辨能力。
Objective To establish a rapid detection method of Vibrio cholerae to improve the speed and detection rate of emergency detection of cholera epidemic and trace the source of suspicious infection. Methods Bacterial isolation and identification, real-time fluorescent PCR rapid detection, colloidal gold immunochromatography rapid test paper method for simultaneous detection of cholera epidemic collected during the collection of patients with water samples, the external environment of water quality, food and aquatic products, and its results Comparative analysis; Vibrio cholerae O139 was isolated by pulsed field gel electrophoresis (PFGE) electrophoresis typing. Results The positive rate of real-time fluorescence PCR and colloidal gold immunochromatography rapid test were higher in the three detection methods. The positive rates of routine bacterial isolation and identification were lower. The three methods had their advantages and disadvantages. PFGE molecular typing analysis suggested that Times O139 cholera outbreak source for the turtle in the aquaculture market, contaminated patients with food consumption. Conclusion Fluorescent PCR and colloidal gold immunochromatography rapid test strip method is an effective supplement to strain isolation and culture methods. PFGE has better discrimination ability for genetic homology analysis of V. cholerae from different sources.