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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 technologies in an anthrax epidemic in Beijing in July 2016 to explore a reasonable and rapid laboratory test method for anthrax epidemic situation. Methods Samples of esophageal mucosa from patients with suspected skin anthrax and smear samples of sera and environmental samples of anthrax epidemic sites were collected. Bacillus anthracis was detected by real-time fluorescence PCR. At the same time, Bacillus anthracis was isolated and identified by bacterial isolation culture method, and the isolated strains were detected by a new generation of sequencing. Results All patients’ smear samples and dead bovine blood-stained soil samples were positive by real-time PCR. One case of anthrax in the affected area of the skin smear samples and dead soil samples of bovine blood plasma were isolated from Bacillus anthracis, belong to the A.Br.001 / 002 group. Serum samples from 5 patients were positive for anthrax capsular antibodies, of which 1 case of double serum was positive for antibodies. Conclusion Laboratory test results fully proved that the outbreak was caused by Bacillus anthracis. Real-time fluorescence PCR, a new generation of molecular biology methods such as sequencing in the rapid identification of anthrax epidemic of great significance.