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[1962 - 2011年上海分离的霍乱弧菌菌株的表型和分子特征鉴定]

[Characterization of phenotype and molecular characteristics on Vibrio cholerae strains isolated in Shanghai, 1962-2011].

作者信息

Tu Li-hong, Chen Min, Li Wei, Zhang Xi, Chen Hong-You, Wang Wen-Jing

机构信息

College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620, China.

Department of Microbiology, Shanghai Municipal Center for Disease Control and Prevention.

出版信息

Zhonghua Liu Xing Bing Xue Za Zhi. 2013 Aug;34(8):815-8.

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 occludens 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 O139 group showed higher prevalence of resistance to several antibiotics compared with O1 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 of ctxA+ 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.

摘要

目的

描述1962年至2011年在上海分离出的霍乱弧菌菌株的表型和分子特征。

方法

采用K-B试验研究霍乱弧菌菌株的抗生素耐药性。应用聚合酶链反应(PCR)检测7种与毒力相关的基因,包括霍乱毒素(ctxA)、小带闭合毒素(zot)、辅助霍乱肠毒素(ace)、溶血素(hlyA)、毒素调节菌毛(tcpA)、外膜蛋白(ompU)和调节蛋白基因(toxR)。通过脉冲场凝胶电泳(PFGE)评估遗传关系,并使用BioNumerics软件对图谱进行聚类。

结果

1962年至1996年分离出的霍乱弧菌菌株对大多数抗生素敏感。然而,2005年至2011年分离出的菌株对多种抗生素耐药。霍乱弧菌O139群对几种抗生素的耐药率高于O1群,O139产毒株的耐药率高于非产毒株。2005年至2011年分离出的大多数O1菌株不产毒,而2005年至2011年分离出的O139菌株几乎都产毒。2005年至2011年从河流中未检测到ctxA+菌株。从水产品中检测到的O1菌株的主要基因类型为hlyA+ toxR+ ompU+,而O139菌株的主要基因类型为hlyA+ toxR+ ompU+ ctxA+ ace+ zot+ tcpA+。使用PFGE,222株霍乱弧菌菌株被分为121种分子类型。O139菌株分为3个簇,O1菌株分为5个簇。

结论

1962年至2011年在上海分离出的霍乱弧菌菌株特征发生了很大变化,提示应更加关注霍乱弧菌菌株抗生素耐药性的增加。

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