副溶血性弧菌恒温荧光法检测及快速检测试剂盒的初步应用  

Real-time isothermal detection of Vibrio parahaemolyticus and its preliminary application in rapid detection kits

作　　者：杨家齐 田明胜 朱智 殷荣永 钟少水 唐皓轩 邵雪阳 倪培恩 王大鹏[4] 张文敏[5] 石育娇 戚成 冯元琦 何更生[1] YANG Jia-qi;TIAN Ming-sheng;ZHU Zhi;YIN Rong-yong;ZHONG Shao-shui;TANG Hao-xuan;SHAO Xue-yang;NI Pei-en;WANG Da-peng;ZHANG Wen-min;SHI Yu-jiao;QI Cheng;FENG Yuan-qi;HE Geng-sheng(School of Public Health,Fudan University,Shanghai 200032,China;Shanghai Municipal Food and Drug Administration,Shanghai 200021,China;Shanghai Gemple Biotechnology Co.,Ltd.,Shanghai 201210,China;School of Agriculture and Bi-ology,Shanghai Jiao Tong University,Shanghai 200240,China;School of Medical Instrument and Food Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China;Guangzhou Deaou Biotechnology Co.,Ltd.,Guangzhou 510663,China)

机构地区：[1]复旦大学公共卫生学院,上海200032 [2]上海食品药品监督管理局,上海200021 [3]上海捷易生物科技有限公司,上海201210 [4]上海交通大学农业与生物学院,上海200240 [5]上海理工大学医疗器械与食品学院,上海200093 [6]广州迪澳生物科技有限公司,广州510663

出　　处：《上海预防医学》2018年第5期386-391,共6页Shanghai Journal of Preventive Medicine

基　　金：上海市科学技术委员会科技支撑项目(13391901402)

摘　　要：【目的】采用恒温荧光PCR方法及快速检测试剂盒对副溶血弧菌的扩增结果进行实时检测,并对其检测限、灵敏度及特异度等进行测试。【方法】通过三个实验阶段(实验室自配阳性样品测试阶段、腹泻病人模拟生物样本测试阶段、市场食品样测试阶段),验证仪器及试剂盒的稳定性、对腹泻样本和市场食品样本检测的可靠性,及其适用范围。【结果】实验室自配样品检测结果较好,检测限达到7~100 cfu/m L;腹泻样和市场食品样的检测假阳性率较高,但灵敏度极好,三个阶段合计灵敏度为99.6%,特异度为55.9%,真阳性率为85.1%,真阴性率为98.3%。【结论】该恒温荧光PCR方法及快速检测试剂盒灵敏度高,稳定性好,适用范围广,在控制实验操作过程及实验条件,提高特异度后,值得进一步推广,对我国食源性副溶血性弧菌的检测具有重要意义。[Objective] To verify the limit of detection as well as sensitivity and specificity of real-time isothermal detection and rapid detection kit of Vibrio parahaemolyticus（ VP）. [Methods]By detecting three groups of samples-laboratory prepared samples,commercial food samples and diarrhea patients samples,the stability and reliability as well as the scope of application of the method and kit were verified. [Results] Laboratory configuration sample detection showed a stable and reliable result,in which the detection limit reached 7 to 100 cfu/m L. Although the FPR of food sample and diarrhea patients sample were relatively high,the sensitivity was great. The total sensitivity,specificity,true positive rate,true negative rate were respectively 99. 6%,55. 9%,85. 1%,98. 3%. [Conclusion]The real-time isothermal detection and rapid detection kit have a great sensitivity for VP detection. Also the results are stable and essentially in agreement with the culture method of GB 4789. 7—2013. The method is worth being promoted in application,under the premise that experimental operating steps and experimental conditions are wellcontrolled in order to improve the specificity.

关 键 词：副溶血性弧菌 恒温荧光PCR 快速检测试剂盒 

分 类 号：R115[医药卫生—公共卫生与预防医学]