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Journal of Clinical Microbiology, March 1998, p. 756-763, Vol. 36, No. 3
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Molecular Analysis of Non-O1, Non-O139 Vibrio cholerae Associated with an Unusual Upsurge in the Incidence of Cholera-Like Disease in Calcutta, India

Charu Sharma,1 M. Thungapathra,1 A. Ghosh,1 Asish K. Mukhopadhyay,2 Arnab Basu,2 Rupak Mitra,2 Indira Basu,2 S. K. Bhattacharya,2 T. Shimada,3 T. Ramamurthy,4 T. Takeda,4 S. Yamasaki,5 Y. Takeda,5 and G. Balakrish Nair2,*

Institute of Microbial Technology, Chandigarh,1 and National Institute of Cholera and Enteric Diseases, Calcutta,2 India, and Laboratory of Enteric Infection 1, National Institute of Infectious Diseases,3 and Research Institute, International Medical Center of Japan,5 Tokyo 162, and Department of Infectious Diseases Research, National Children's Medical Research Center, Tokyo 154,4 Japan

Received 17 June 1997/Returned for modification 22 October 1997/Accepted 15 December 1997

There was an inexplicable upsurge in the incidence of non-O1, non-O139 Vibrio cholerae among hospitalized patients admitted to the Infectious Diseases Hospital, Calcutta, India, between February and March 1996. Of the 18 strains of V. cholerae isolated during this period, 15 belonged to the non-O1, non-O139 serogroups (4 belonged to O144, 3 belonged to O11, 1 each belonged to O6, O8, O12, O19, O39, and O58, and 2 strains could not be typed), 2 belonged to the O139 serogroup, and 1 belonged to the O1 serogroup. Cell-free culture supernatants of 13 representative non-O1, non-O139 V. cholerae strains evoked a distinct cytotoxic effect on CHO and HeLa cells, and the strains examined produced the nonmembrane-damaging cytotoxin. By several PCR assays, it was determined that none of the non-O1, non-O139 strains were positive for the ctxA, zot, ace, and tcpA genes and for the genes representing the heat-labile toxin, heat-stable toxin, and verotoxin of Escherichia coli and the various variants of these genes. Studies on the clonality of non-O1, non-O139 V. cholerae strains by restriction fragment length polymorphism (RFLP) analysis of rRNA genes and of other genes (hlyA, hlyU, hlx, toxR, and attRS1) and by pulsed-field gel electrophoresis (PFGE) collectively indicate that the upsurge which occurred in February and March 1996 was caused by strains belonging to different clones. Overall, there was an excellent correlation between the results of ribotyping, RFLP analysis of various genes, and PFGE, with strains belonging to a particular serogroup showing nearly identical restriction patterns and PFGE profiles. It is clear from this study that some serogroups of V. cholerae can cause diarrhea by a mechanism quite different from that of toxigenic V. cholerae O1 and O139, and we have proposed the nomenclature of enteropathogenic V. cholerae to include these serogroups.

* Corresponding author. Mailing address: National Institute of Cholera and Enteric Diseases, P-33, CIT Road, Scheme XM, Beliaghata, Calcutta 700 010, India. Phone: 91-33-3504598. Fax: 91-33-3505066. E-mail: krishgb{at}giascl01.vsnl.net.in.

Journal of Clinical Microbiology, March 1998, p. 756-763, Vol. 36, No. 3
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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