一起炭疽疫情中PCR和新一代测序技术应用研究

Application of PCR and next generation sequencing in an anthrax epidemic

目的对2016年7月北京市发生的一起炭疽疫情中PCR和新一代测序技术的应用情况进行描述和分析,探讨合理和快速的炭疽疫情实验室检测方法。方法采集疑似皮肤炭疽患者的焦痂处涂抹样本和血清样本及炭疽疫情现场环境样本,采用实时荧光PCR法检测炭疽芽胞杆菌。同时采用细菌分离培养法分离和鉴定炭疽芽胞杆菌,应用新一代测序法对分离到的菌株进行检测。结果所有患者的涂抹样本和病死牛血污染土壤样本实时荧光PCR法检测为阳性。1例炭疽患者患处皮肤涂抹样本和病死牛血污染的土壤样本中分别分离到炭疽芽胞杆菌,同属于A.Br.001/002组。5例患者血清中炭疽荚膜抗体阳性,其中1例双份血清检测到抗体阳转。结论实验室检测结果充分证明了此次疫情是由炭疽芽胞杆菌感染引起。实时荧光PCR、新一代测序等分子生物学方法在快速鉴定炭疽疫情中具有重要意义。

Objective To describe and analyze the application of PCR and next generation sequencing in the laboratory testing of an anthrax epidemic in Beijing in July 2016,so as to investigate the rational and rapid laboratory methods for anthrax epidemic. Methods Swabbing samples on the typical black eschars and serum samples of the cutaneous anthrax patients as well as environmental samples were collected. Rapid testing methods,including real time PCR and traditional bacterial analysis,were performed to detect Bacillus Anthracis. Results All swabbing samples and soil sample contaminated with blood of a dead cow were positive with real time PCR. Two stains of B. anthracis were isolated and confirmed from the sample of one patient and soil sample contaminated with blood of the dead cow. Both stains belonged to A. Br. 001/002 group. 5 serum samples were positive of antibodies against capsule of B. anthracis. Serum conversion was observed in the paired serums from one patient. Conclusion Multiple laboratory results proved that the epidemic was caused by B. anthracis infections. Real time PCR,next generation sequencing should play more important role in the laboratory analysis of anthrax epidemics.