口岸入境人员腹泻病原体快速检测流程的建立

A quick detection process for diarrhea pathogens among entry population at frontier ports

目的建立一种适合国境口岸入境人员腹泻样本中病原体快速检测的流程。方法联合使用荧光PCR和细菌培养分离鉴定法,先用荧光PCR法对引起人类腹泻的常见4种病毒(轮状病毒、诺如病毒、星状病毒、札如病毒)和11种细菌(沙门菌、志贺菌、肠致病性大肠埃希菌、肠出血性大肠埃希菌、肠侵袭性大肠埃希菌、肠产毒性大肠埃希菌、霍乱弧菌、副溶血弧菌、李斯特菌、空肠弯曲菌、金黄色葡萄球菌)进行核酸检测,阳性样本再采用选择性培养基进行有针对性的细菌培养分离鉴定。结果 2016年8-10月,用建立的检测流程从14份口岸入境人员肛拭子样本中检出2份肠致病性大肠埃希菌、1份肠炎沙门菌、1份温和气单胞菌和1份铜绿假单胞菌,检测时间均只用3 d。结论建立的荧光PCR结合细菌培养分离鉴定的联合检测流程,可大幅缩短检测时间,值得在口岸推广应用。

Objective To establish a quick detection process for diarrhea pathogens among entry population at frontier ports. Methods The quick detection process combined fluorescent Real-time PCR and bacterial culture method.Firstly,fluorescent Real-time PCR was applied for the detection of four common diarrhea viruses（rotavirus,norovirus,astrovirus and sapovirus） and eleven bacteria,including Salmonella,Shigella,Enteropathogenic Escherichia coli（EPEC）,Enterohemorrhagic Escherichia coli（EHEC）,Enteroinvasive Escherichia coli（EIEC）,Enterotoxigenic Escherichia coli（ETEC）,Vibrio cholera,Vibrio parahaernolyt icus,Listeria monocytogenes,Campylobacter jejuni and Staphylococcus aureus. Then target pathogens were detected by selective culture method. Results A total of 14 anal swab specimens collected during August to October,2016,were detected by this quick detection process. Two EPEC strains,one Salmonella enteritidis strain one Aeromonas sobria strain and one Pseudomonas aeruginosa strain had been identified.This detection process was completed in three days. Conclusion The quick detection process could not only save time but also increase the positive rate,and it deserves to be promoted widely at frontier ports.