| 屠丽红,陈敏,李伟,张曦,陈洪友,王文静.上海市1962-2011年霍乱弧菌表型特征及分子分型研究[J].Chinese journal of Epidemiology,2013,34(8):815-818 |
| 上海市1962-2011年霍乱弧菌表型特征及分子分型研究 |
| Characterization of phenotype and molecular characteristics on Vibrio cholerae strains isolated in Shanghai, 1962-2011 |
| Received:April 22, 2013 |
| DOI： |
| KeyWord: 霍乱弧菌|表型|分子分型|脉冲场凝胶电泳 |
| English Key Word: Vibrio cholerae|Phenotype|Molecular type|Pulsed-field gel electrophoresis |
| FundProject:新-轮上海市卫生系统优秀学科带头人计划(XBR20l1051);上海市公共卫生重点学科卫生微生物学(12GWZX0801) |
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| Abstract: |
| 目的 分析1962-2011年上海市霍乱弧菌的表型及分子分型特征.方法 采用WHO推荐的改良K-B纸片法,对222株霍乱弧菌进行11种抗菌药物(头孢曲松、强力霉素、诺氟沙星、环丙沙星、复方新诺明、丁胺卡那霉素、四环素、氯霉素、萘啶酸、氨苄西林、庆大霉素)敏感试验.以PCR检测霍乱毒素基因(ctxA)、小带联结毒素基因(zot)、辅助霍乱肠毒素基因(ace)、溶血素基因(hlyA)、毒素协调菌毛基因(tcpA)、外膜蛋白基因(ompU)和调控蛋白基因(toxR).采用脉冲场凝胶电泳(PFGE)方法对菌株进行分子分型,用BioNumerics软件分析电泳图谱.结果 222株霍乱弧菌经药物敏感试验分析显示,1962-1996年的菌株对多种药物敏感,2005-2011年的菌株对多种药物耐药.O139群耐药率明显高于O1群,O139群产毒株的耐药率比非产毒株高.毒力基因分析显示,1962-1996年霍乱患者来源菌株多为O1群产毒株,2005-2011年患者来源菌株多为O139群产毒株,水体来源菌株未检出ctxA基因,O1群水产来源菌株以hlyA+ toxR+ ompU+为主,占25.6％(11/43),O139群水产来源菌株以hlyA+ toxR+ompU+ctxA +ace +zot+ tcpA+为主,占76.1％(16/21).PFGE分析将222株菌分为121个PFGE型,O139群分为3个聚类,O1群分为5个聚类.结论 上海市霍乱弧菌随着时间推移表型及分子特征均发生了很大变化,耐药情况加重. |
| English Abstract: |
| Objective To describe the phenotype and molecular characteristics of Vibrio (V.) cholerae strains isolated in Shanghai,from 1962 to 2011.Methods K-B test was used to investigate the antibiotic resistance of V.cholerae strains.PCR was applied to detect seven virulence-related genes including cholera toxin (ctxA),zonula oecludens toxin (zot),accessory cholera enterotoxin (ace),hemolysin (hlyA),toxin-coregulated pilus (tcpA) outer membrane protein (ompU) and the regulatory protein genes (toxR).Genetic relation was assessed by pulsed-field gel electrophoresis (PFGE) and the patterns were clustered by BioNumerics software.Results V.cholerae strains isolated from 1962 to 1996 were sensitive to most of the antibiotics.However,the strains isolated from 2005 to 2011 were resistant to many antibiotics.V.cholerae O 139 group showed higher prevalence of resistance to several antibiotics compared with O l group,and the resistance rate of the O139 toxigenic isolates was higher than that of the non-toxigenic isolates.Most of the O1 strains isolated from 2005 to 2011 were non-toxigenic while O139 strains isolated from 2005 to 2011 were almost toxigenic.There were no strains ofctxA+ detected from the rivers from 2005 to 2011.Main gene type of the O1 strains detected from the aquatic products was hlyA+toxR+ompU+,while that of the O139 strains was hlyA+toxR+ompU+ ctxA + ace +zot + tcpA +.Using PFGE,222 V.cholerae strains were subtyped into 121 molecular types.O139 strains were divided to three clusters and O1 strains to five clusters.Conclusion The characteristics of V.cholerae strains isolated in Shanghai from 1962 to 2011 showed great changes,suggesting that more attention should be paid to the multiplication on antibiotic resistance of V.cholerae strains. |
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