46株肺炎链球菌的血清型别和毒力因子基因的保守性分析  

Analysis for serotypes of 46 strains of invasive Streptococcus pneumoniae and their conservation of virulence factors

作　　者：刘孟涓 张群[2] 王虹[1] 胥文春[1] 何於娟[1] 尹一兵[1] 张雪梅[1] 

机构地区：[1]重庆医科大学检验医学院,重庆400016 [2]重庆医科大学附属儿童医院检验科,重庆400014

出　　处：《临床检验杂志》2013年第12期934-937,共4页Chinese Journal of Clinical Laboratory Science

基　　金：重庆市卫生局医学科研项目面上项目(2011-2-247)

摘　　要：目的研究临床分离的肺炎链球菌菌株的血清型和重要毒力因子碱基序列的保守情况，为临床肺炎链球菌疫苗的使用和蛋白质疫苗的研发提供参考依据。方法收集2010—2013年重庆医科大学附属儿童医院11岁以下儿童中分离的46株肺炎链球茵，用多重PCR进行血清型分型。同时PCR扩增肺炎链球菌的11个毒力基因ply、lytA、nanA,phtD、c如P、cps2A、bacterio—cin、psaA,piaA、pcsB和stkP，测序后分析碱基序列的突变情况。结果46株肺炎链球茵共检出13种血清型，其中19A（19．57％）为最常见的血清型，其次为23F和19F（各占15．22％）。临床菌株毒力基因ply、lytA、nanA、phtD、czpP、cps2A、bacteriocin,psaA,piaA、pcsB和stkP的碱基序列保守性分别为97．82％、96．31％、82．96％、87．50％、98．48％、83．68％、66．27％、99．46％、98．63％、88．72％和96．67％。结论在重庆地区儿童感染的肺炎链球菌中，血清型19A分布比例最高，本地区儿童接种肺炎链球菌疫苗时应首选含有19A血清型荚膜多糖抗原的PCVl3疫苗。毒力因子ply、stkP、piaA、lytA、cop和psaA基因在临床菌株中保守性较高，具有作为肺炎链球菌疫苗候选蛋白质的潜力。Objective To investigate the serotypes of the clinical strains of Streptococcus pneumoniae （ S. pn） and the conservatism of the base sequences of the important virulence factors in order to provide information for the application of pneumococcal conjugate vac- cine （PCV） and the development of protein vaccine. Method A total of 46 pneumococci isolates were collected from the children bel- low 11 years of age in the Children＇s Hospital of Chongqing Medical University between 2010 and 2013. Multiplex PCR typing was used for serotyping of the isolates. The 11 virulence genes, i.e. , ply, lytA, nanA, phtD, clpP, cps2A, bacteriocin, psaA, piaA, pcsB and stkP were also amplified by PCR and the PCR products sequencing was performed to analyze the mutation of base sequences. Results Among the 46 isolates 13 serotypes were identified. The most prevalent serotype was 19A （ 19.57% ） followed by 23F and 19F （both 17.02% ）. The conservatism of the virulence genes ply, lytA ,nanA, phtD, clpP, cps2A, bacteriocin, psaA, piaA, pcsB and stkP were as following in order： 97.82%, 96.31%, 82.96%, 87.50%, 98.48%, 83.68%, 66.27%, 99.46%, 98.63%, 88.72% and 96.67%. Conclusion The proportion of serotype 19A was the highest among the invasive S. pn in Chongqing, so the children should accept vaccination of PCV13, in which the antibody to capsular polysaccharide of serotype 19A was contained, to protect from S. pn in- fection. In the clinical isolates of S. pn, the virulence factors, ply, stkP, piaA, lytA, clpP and psaA showed high conservation, which may have the potential to become the candidate protein for pneumococcal vaccine.

关 键 词：肺炎链球菌 血清分型 毒力因子 

分 类 号：R515[医药卫生—内科学] R378.12[医药卫生—临床医学]