一起产气荚膜梭菌食物中毒事件的实验室检测分析

冀国强; 李颖; 张爽; 吕金昌; 张赫; 马红梅; 逄波; 张茂俊

doi:10.3784/j.issn.1003-9961.2020.06.019

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

摘要

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1064 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。

Abstract:
ObjectiveTo identify the pathogen of one food poisoning event using by three detection methods.
MethodReal-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 1064 Copies/μl. Twenty two C. perfringens isolates had 2 different PFGE patterns.
ConclusionLaboratory analysis through three detection methods indicated that this food poisoning might be caused by C. perfringens.

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