检测肺炎球菌疫苗功能性抗体的标准株分子特征分析  

Molecular genetic analysis of standard strains for functional antibody detection of pneumococcal polysaccharide vaccine

作　　者：黄洋[1] 徐潇[1] 李康[1] 李江姣[1] 石继春[1] 郑锐 孙文媛 叶强[1] Huang Yang;Xu Xiao;Li Kang;Li Jiangjiao;Shi Jichun;Zheng Rui;Sun Wenyuan;Ye Qiang(National Center for Medical Culture Collections,National Institutes for Food and Drug Control,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院中国医学细菌保藏管理中心,北京102629

出　　处：《中华实验和临床病毒学杂志》2021年第5期509-513,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：国家科技资源共享服务平台(国家病原微生物资源库-NPRC-32)。

摘　　要：目的检测肺炎球菌疫苗功能性抗体的标准株分子特征分析,为肺炎球菌疫苗的临床评价奠定基础。方法对24株检测肺炎球菌疫苗功能性抗体的标准株的16S rRNA基因片段进行PCR扩增、测序,并运用多位点序列分析(MLST)和脉冲场凝胶电泳(PFGE)等方法,对相关菌株进行分子分型。结果16S rRNA基因序列比对结果显示24株菌株的基因序列与肺炎链球菌模式株NCTC 7465的16S rRNA基因序列(LN831051)的相似性在98.35%~99.86%之间,碱基的差异在2~23 bp;多位点序列分析结果显示24株标准菌株的ST型各不相同;PFGE分析结果表明24株标准菌株100~1000 Kb大小的DNA片段得到了较好的分离效果,各血清群/型的菌株各具其特征性的条带(12~15个片段)。结论本研究发现检测肺炎球菌疫苗功能性抗体的标准株的ST型和PFGE图型均不相同,具有代表性,为肺炎球菌功能性抗体检测用标准菌株质量控制方法提供参考。Objective To analyze molecular genetic characteristics of standard strains for functional antibody detection of pneumococcal polysaccharide vaccine,and provide the foundation for clinical research of pneumococcal polysaccharide vaccine.Methods The 24 strains for functional antibody detection of pneumococcal polysaccharide vaccine were detected by PCR amplification and sequencing using standard strain 16S rRNA gene fragment,and the molecular typing of the related strains was performed by multi locus sequence analysis(MLST)and pulsed field gel electrophoresis(PFGE).Results The result of 16S rRNA gene sequence alignment showed that the similarity between the gene sequences of the 24 strains and that of NCTC 7465(LN831051)ranged from 98.35%to 99.86%,and the base difference was 2-23 bp.Multi locus sequence analysis showed that the ST types of 24 standard strains were different.PFGE analysis showed that 24 standard strains had good isolation effect from DNA fragments of 100 kb-1000 kb,and each serotype had its own characteristic bands(12-15 fragments).Conclusions This study analyzed the molecular genetic characteristics of the standard strains for the detection of pneumococcal functional antibodies for the first time,and may provide a reference for the quality control method of the standard strains for the subsequent detection of pneumococcal functional antibodies.

关 键 词：肺炎球菌疫苗 功能抗体检测 标准菌株 分子分型 

分 类 号：R392-33[医药卫生—免疫学]