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Journal of Clinical Microbiology, June 2003, p. 2306-2310, Vol. 41, No. 6
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.6.2306-2310.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Epidemiology of Vibrio cholerae O139 in China: Polymorphism of Ribotypes and CTX Elements

Mei Qu,{dagger} Jing Xu, Yanpeng Ding,{ddagger} Ruibai Wang, Peng Liu, Biao Kan,* Guoming Qi, Yanqing Liu, and Shouyi Gao

Priority Laboratory of Medical Molecular Bacteriology of Ministry of Health, Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Beijing, People's Republic of China

Received 11 June 2002/ Returned for modification 10 October 2002/ Accepted 10 March 2003

Vibrio cholerae O139, the second etiological serogroup of cholera, triggered the first outbreak of O139 cholera in China in 1993. To analyze the clone polymorphism of O139 isolates in China, 117 strains of V. cholerae O139, isolated from different areas in China between 1993 and 1999, were selected to characterize the phylogenetic relationships by molecular techniques. Analysis of restriction fragment length polymorphism in the conserved 16S rRNA gene revealed seven different ribotypes within the 117 strains. Among these strains, there were eight that lacked the cholera toxin gene (ctxAB), zot, and the repetitive sequence (RS); these eight strains belonged to three individual ribotypes. Our results suggested that V. cholerae O139 strains in China had clone diversity in phylogeny. The results of our hybridization patterns for CTX genetic elements (ctxAB, zot, and RS) showed that CTX{Phi} genomes in most V. cholerae O139 strains had two or more copies and had extensive restriction patterns even for the strains which belong to the same ribotype. For 22 (20.1%) strains, the copies of ctxAB were different from those of zot, suggesting that a ctxAB-negative CTX{Phi} genome may exist in O139 strains. This ctxAB-negative CTX{Phi} genome may coexist with the intact CTX{Phi} genome in a strain. In addition, the dendrogram for I-CeuI-generated pulsed-field gel electrophoresis patterns showed that V. cholerae serogroup O139 has a closer relationship with one strain of serogroup O22 than with the strains of serogroup O1. The results of this study showed the clonal diversity and the distribution of O139 strains in China, suggesting multiple origins of the O139 cholera epidemic or sporadic events.

* Corresponding author. Mailing address: Priority Laboratory of Medical Molecular Bacteriology of Ministry of Health, Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, P.O. Box 5, Changping, Beijing, 102206, People's Republic of China. Phone: 8610-61739458. Fax: 8610-61730233. E-mail: kanb{at}btamail.net.cn.

{dagger} Present address: National Reference Laboratory on Reproductive Health Research of Ministry of Health, Institute of Reproductive and Child Health of Peking University, Beijing, 100083, People's Republic of China.

{ddagger} Present address: Division of Geographic Medicine and Infectious Diseases, Howard Hughes Medical Institute, New England Medical Center, Boston, MA 02111.

Journal of Clinical Microbiology, June 2003, p. 2306-2310, Vol. 41, No. 6
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.6.2306-2310.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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