一起产气荚膜梭菌食物中毒事件的实验室检测分析  

Laboratory analysis on one food poisoning event caused by Clostridium perfringens

作　　者：冀国强[1] 李颖[1,2] 张爽[1,2] 吕金昌[1] 张赫 马红梅[1,2] 逄波 张茂俊[2,3] Ji Guoqiang;Li Ying;Zhang Shuang;Lyu Jinchang;Zhang He;Ma Hongmei;Pang Bo;Zhang Maojun(Shunyi District Center for Disease Control and Prevention,Beijing 101300,China;Workstation for Microbial Infectious Disease,Shunyi District Center for Disease Control and Prevention,Beijing 101300,China;National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]北京市顺义区疾病预防控制中心,北京101300 [2]北京市顺义区疾病预防控制中心微生物感染性疾病检测工作站,北京101300 [3]中国疾病预防控制中心传染病预防控制所,北京102206

出　　处：《疾病监测》2020年第6期547-551,共5页Disease Surveillance

基　　金：国家科技重大专项(No.2018ZX10712–001)。

摘　　要：目的应用3种方法对一起食物中毒事件中的可疑样品进行检测和结果分析。方法应用实时荧光PCR、数字PCR和平板计数培养法检测食物中毒事件中的可疑食品盐水猪肝,并对分离菌株用脉冲场凝胶电泳(PFGE)进行分型。应用细菌生化试验、实时荧光PCR鉴定可疑食品中的产气荚膜梭菌;应用数字PCR对毒力基因进行绝对定量。结果可疑食品产气荚膜梭菌平板计数值为4 400 000 CFU/g,并分离到22个产气荚膜梭菌单菌落,实时荧光PCR检测可疑食品样本产气荚膜梭菌plc和cpe基因阳性,其中3个菌落plc+/cpe+,19个菌落plc+/cpe-;数字PCR检测可疑食品样本plc基因绝对定量值为1 064拷贝/μl;22个单菌落分为2种PFGE带型。结论通过3种方法检测可疑食品中产气荚膜梭菌,说明该起食物中毒可能由产气荚膜梭菌导致。Objective To identify the pathogen of one food poisoning event using by three detection methods. Method Real-time PCR, digital PCR and plate count culture were applied in the detection of suspected contaminated food samples(salt water pig liver);Pulsed-field gel electrophoresis was performed for the typing of the isolates. Biochemical bacteria identification and real-time PCR were performed for the Clostridium perfringens isolates from the suspected contaminated food samples;digital PCR was performed for the absolute quantification of virulence genes. Results Plate count culture value of C. perfringens was 4 400 000 CFU/g for the suspected contaminated food samples, and 22 C. perfringens isolates were obtained, which were positive for both plc gene and cpe gene of C. perfringens indicated by real-time PCR;and 3 isolates were plc+/cpe+, 19 isolates were plc+/cpe-. Absolute quantification value of plc gene was 1 064 Copies/μl. Twenty two C.perfringens isolates had 2 different PFGE patterns. Conclusion Laboratory analysis through three detection methods indicated that this food poisoning might be caused by C. perfringens.

关 键 词：产气荚膜梭菌 食物中毒 实时荧光PCR 数字PCR 平板计数法 

分 类 号：R155.3[医药卫生—营养与食品卫生学]