2012—2019年北京市朝阳区霍乱弧菌毒力基因和 PFGE分子分型特征分析  

Serotypes, virulence genes and PFGE profiles of Vibrio cholerae strains isolated in Chaoyang district of Beijing, 2012—2019

作　　者：赵剑虹[1] 顾琳[1] 高艳[1] 焦洋[1] 曲梅[2] 严寒秋[2] ZHAO Jian-hong;GU Lin;GAO Yan;JIAO Yang;QU Mei;YAN Han-qiu(Beijing Chaoyang District Center for Disease Control and Prevention,Beijing 100021,China;不详)

机构地区：[1]北京市朝阳区疾病预防控制中心,北京100021 [2]北京市疾病预防控制中心北京市预防医学研究中心食物中毒诊断溯源技术北京市重点实验室

出　　处：《中国预防医学杂志》2021年第12期903-908,共6页Chinese Preventive Medicine

基　　金：国家科技重大专项(2018ZX10714002-003-002)。

摘　　要：目的了解2012—2019年北京市朝阳区霍乱弧菌毒力基因和脉冲场凝胶电泳(pulsed field gel electrophoresis, PFGE)分子分型特征。方法收集2012—2019年北京市朝阳区分离的可疑O1群和O139群霍乱弧菌菌株,常规方法鉴定后,用实时荧光定量PCR(real-time PCR)对菌株进行分子水平鉴定并检测霍乱弧菌ctxAB基因;用PCR方法检测toxR、hlyA-El、hlyA-Cl、ompU、tcpA-El、tcpA-Cl、tcpI、rtxC、zot和ace 10种毒力基因。用NotⅠ和SfiⅠ两种内切酶对菌株进行PFGE分子分型试验,用BioNumerics(Version 5.01,Applied Maths, Inc)软件处理和读取PFGE图像,进行菌株图谱相似度聚类分析。结果共分离到37株霍乱弧菌,O1小川型/产毒株、O1小川型/非产毒株、O1稻叶型/非产毒株和O139/产毒株构成比分别为2.70%(1/37)、67.57%(25/37)、27.03%(10/37)和2.70%(1/37)。所有菌株toxR、hlyA-El、hlyA-Cl和rtxC基因携带率为100.00%,其他毒力基因ompU、tcpA-El、tcpA-Cl、tcpI、zot、ace和ctxAB的携带率分别为56.76%(21/37)、45.95%(17/37)、43.24%(16/37)、54.05%(20/37)、24.32%(9/37)、24.32%(9/37)和5.41%(2/37);37株霍乱弧菌的11种毒力基因分为携带谱P1~P22,携带毒力基因为4~10种,携带5种毒力基因的P3占优势(构成比为21.62%,8/37)。26株O1群小川型PFGE图谱相似度为62.90%~100.00%,型别为CYXC01~CYXC20;10株O1稻叶型PFGE图谱相似度为67.90%~100.00%,型别为CYDY01~CYDY09。36株O1群霍乱弧菌的21种毒力基因携带谱与29个PFGE型别相互对应分散。结论 2012—2019年北京市朝阳区霍乱弧菌以O1小川型/非产毒株为主,毒力基因携带谱和PFGE型别多样,同种毒力基因携带谱与同一PFGE型别相互对应不完全一致。Objective To characterize the serotypes, virulence genes and the profiles of pulsed field gel electrophoresis(PFGE) of Vibrio cholerae strains isolated in Chaoyang district of Beijing from 2012 to 2019. Methods Suspicious strains of Vibrio cholerae O1 and O139 isolated in Chaoyang district in Beijing from 2012 to 2019 were collected and confirmed by conventional methods. Real-time PCR was used for the phenotypic determination of O1/O139 Vibrio cholerae strains and the detection of ctxAB gene. Other virulence genes as toxR, hlyA-El, hlyA-Cl,ompU, tcpA-El, tcpA-Cl, tcpI,RtxC, zot and ace 10 were determined by conventional PCR. Two endonucleases, Not I and Sfi I, were used for molecular genotyping of V. Cholerae strains carried out by PFGE, and the clustering of the similarity of the strain maps were analyzed using Bio Numerics software. Results From 2012 to 2019,a total of 37 strains of V.cholerae were isolated in Chaoyang district in Beijing,including 1 strain of O1 Ogawa toxigenic V.Cholerae(2.70%),25 strains of O1 Ogawa non-toxigenic V.Cholerae(67.57%),10 strains of O1 Inaba non-toxigenic V.Cholerae(27.03%)and O1 strain of O139 toxigenic V.Cholerae(2.70%).All strains carried toxR,hlyA-El,hlyA-Cl and rtxCgenes,while virolence genes of ompU,tcpA-El,tcpA-Cl,tcpI,zot,ace and ctxAB were identified in56.76%(21/37),45.95%(17/37),43.24%(16/37),54.05%(20/37),24.32%(9/37),24.32%(9/37)and 5.41%(2/37)of isolates,respectively.37 strains of Vibrio cholera were divided into 11 virulence gene carrying profiles of P1-P22,having 4-10 virulence genes,and P3 with 5 virulence genes was dominant(8/37).PFGE profile showed a 62.90%-100.00% similarity among 26 strains of O1 Ogawa serotype with the genotypes ranging from CYXC01 to CYXC20.Among 10 strains of O1 Inaba serotype,the similarity of the PFGE profile was from 67.90%to100.00%,with the genotypes ranging from CYDY01 to CYDY09.The 21 virulence gene carrying profiles and 29 PFGE genotype patterns of 36 O1 Vibrio cholerae strains were scattered correspondingly. Conclusions From 2012 t

关 键 词：霍乱弧菌 毒力基因 脉冲场凝胶电泳 携带谱 型别 

分 类 号：R516.5[医药卫生—内科学]