肺炎链球菌国家标准株的建立与评价  

Establishment and evaluation of national standard strains of Streptococcus pneumoniae

作　　者：白霜[1] 张鹏[1] 赵伟[1] 郑群[2] 吴疆[1] Bai Shuang;Zhang Peng;Zhao Wei;Zheng Qun;Wu Jiang(Immunization Department,Beijing Center for Disease Prevention and Control,Beijing 100013,China;School of Basic Medicine Sciences,Capital Medical University,Beijing 100069,China)

机构地区：[1]北京市疾病预防控制中心免疫预防所,北京100013 [2]首都医科大学基础医学院,北京100069

出　　处：《疾病监测》2023年第12期1491-1497,共7页Disease Surveillance

基　　金：北京市属医学科研院所公益发展改革试点项目(No.京医研2018-1)。

摘　　要：目的依据《病原微生物菌(毒)种国家标准菌株评价技术标准》(WS/T 812-2022)对肺炎链球菌临床分离株进行筛选,建立肺炎链球菌国家标准株候选株。方法通过对肺炎链球菌候选株菌落形态、染色特征、生化特性等进行菌种确认;通过乳胶凝集实验、荚膜肿胀实验、多重聚合酶链式反应、cpsB基因测序等进行菌种分型鉴定;利用Nanopore三代测序平台对菌种进行全基因组测序。结果在700余株肺炎链球菌临床分离株中,共筛选出21株具有肺炎链球菌典型特征的菌株纳入国家标准株候选株。21株肺炎链球菌经3种方法分型诊断一致证实为血清型3、4、6A、6B、7F、8、9N、9V、10A、11A、12F、14、15B、17F、18C、19A、19F、20、22F、23F、33F。菌株稳定性实验结果显示,连续传代30代后,其菌落形态、染色特性、生化特性以及血清型与原代菌株均保持良好一致性。研究所选21株肺炎链球菌均符合国家标准菌株的要求。结论本研究所获21株肺炎链球菌菌株具有相应病原菌的典型特征,可作为肺炎链球菌相关疾病监测、疫苗评价等工作的国家标准株的候选株。Objective To establish candidates of national standard strains of Streptococcus pneumoniae by screening clinical isolates of S.pneumoniae according to the Technical Requirements for Evaluation of National Standard Strains of Pathogenic Microorganisms(WS/T 812-2022).Methods The S.pneumoniae candidate strains were identified by colony morphology,Gram staining,biochemical characteristics.The serotypes of strains were clarified through Quellung and latex agglutination,multiplex polymerase chain reaction,and cpsB gene sequencing,the whole-genome sequencing was carried out using the Nanopore sequencing platform.Results A total of 21 strains with typical characteristics of S.pneumoniae were screened as the candidate strains among more than 700 clinical isolates of S.pneumoniae.All the 21 strains were consistently confirmed as serotypes 3,4,6A,6B,7F,8,9N,9V,10A,11A,12F,14,15B,17F,18C,19A,19F,20,22F,23F,33F by methods mentioned above of typing.The results of stability experiments showed that the colony morphology,Gram staining,biochemical characteristics,and serotypes maintained good consistency with those of the original strain after consecutively regenerating for 30 generations.The 21 strains of S.pneumoniae selected in this study met all the requirements of the national standard bacterial candidates.Conclusion The 21 strains of S.pneumoniae obtained in this study have the typical characteristics of the corresponding bacteria and can be used as candidates for national standard strains in the surveillance of S.pneumoniae-related diseases and vaccine evaluation.

关 键 词：肺炎链球菌 国家标准株 血清学分型 

分 类 号：R211[医药卫生—中医学] R37[生物学—微生物学] Q939.99