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目的通过采用实时荧光PCR法和培养法对竞赛盲样样品中食源性致病菌的分离鉴定,建立了针对多项目质控样品检测的快速准确方法。方法 2014年江苏省疾病预防控制中心组织的传染病防治技能竞赛中给出的盲样考核样品,依据提供的案例A和B传染病发病临床表现,锁定检测范围,先运用实时荧光PCR法进行致病菌初筛,再以荧光PCR结果为指导,用培养法进行进一步分离鉴定,最终得到正确的检测结果。结果项目A的检测结果为A-049和A-086检出福氏2b型志贺菌,A-093未检出致病菌。项目B检测结果为B-056检出肠出血性大肠杆菌O157:H7,B-022和B-086未检出病原菌。结论荧光PCR对传染病突发事件中致病菌的快速初筛具有重要意义,与培养法结合应用于多项目质控样本检测可减少传统培养方法漏检的可能性,提高检测效率和正确率。
OBJECTIVE: To establish a rapid and accurate method for the quality control of multi-project samples by using real-time fluorescence PCR and culture methods to isolate and identify food-borne pathogens in blind samples. Methods Blind sample samples given in 2014 in Jiangsu Province CDC were selected according to the clinical manifestations of cases A and B and the scope of detection was locked. Real-time PCR was used to detect Pathogenic bacteria screening, and then fluorescence PCR results as a guide, further isolation and identification with the culture method, and ultimately get the correct test results. Results The detection results of item A were Shigella flexneri 2b type A-049 and A-086, but no pathogens were found in A-093. Project B test results for the detection of enteric hemorrhagic Escherichia coli O157: H7 B-056, B-022 and B-086 were not detected pathogens. Conclusion Fluorescent PCR is of great importance to rapid screening of pathogens in emergencies of infectious diseases. Combined with culture method, the detection of multiple quality control samples can reduce the possibility of missed detection by traditional culture methods and improve the detection efficiency and accuracy .