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Journal of Clinical Microbiology, December 1998, p. 3595-3600, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Rapid Diagnosis of Cholera Caused by Vibrio cholerae O139

Wanpen Chaicumpa,1,* Potjanee Srimanote,1 Yuwaporn Sakolvaree,1 Thareerat Kalampaheti,1 Manas Chongsa-Nguan,1 Pramuan Tapchaisri,1 Boonchuay Eampokalap,2 Pikul Moolasart,2 G. Balakrish Nair,3 and Peter Echeverria4

Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University,1 Armed Forces Research Institute of Medical Sciences (AFRIMS),4 Bangkok 10400, and Bamrasnaradura Infectious Diseases Hospital, Nonthaburi Province,2 Thailand, and National Institute of Cholera and Enteric Diseases, Calcutta, India3

Received 13 July 1998/Returned for modification 18 August 1998/Accepted 24 September 1998

Hybridomas secreting specific monoclonal antibodies (MAbs) to Vibrio cholerae serogroup O139 were produced. Six monoclones (hybridomas) secreting MAbs specific only to lipopolysaccharide of V. cholerae O139 strains and which did not cross-react to 137 strains of other enteric microorganisms were obtained. These clones were designated 12F5-G11, 12F5-G2, 15F5-H5, 5B9-F8, 14C9-D2, and 6D2-D8. The immunoglobulin (Ig) heavy chain isotypes secreted by these clones were IgG2b, IgG2b, IgG2b, IgM, IgG2b, and IgG3, respectively. Clone 12F5-G11 was selected for mass production of MAb, which was used as a detection reagent in the antigen detection assay for diagnosis of cholera caused by V. cholerae O139, and this assay was compared to the conventional bacterial isolation method. Five batches of rectal swab cultures in alkaline-peptone water were collected from 6,497 patients with watery diarrhea. These were 6,310 patients admitted to Bamrasnaradura Infectious Diseases Hospital, 16 patients from Krung Thon Hospital, 78 patients from Bangkok Children's Hospital, 19 patients from Karen refugee camps, and 74 Indian patients from the National Institute of Cholera and Enteric Diseases, Calcutta, India. The V. cholerae O139 isolations from the rectal swab cultures and the antigen detection assays (i.e., the MAb-based dot-blot ELISA) were performed by different persons of different laboratories, and the results were revealed after all specimens had been tested. Of the 6,497 samples tested, the dot-blot ELISA correctly identified 42 of 42 V. cholerae O139-positive samples and gave a result of positive for three samples which were culture negative for V. cholerae O139. The diagnostic sensitivity, specificity, and efficacy of the dot-blot ELISA were 100, 99.95, and 99.26%, respectively. The ELISA is easy to perform and relatively inexpensive. It can test multiple samples at a single time, does not require special equipment, and does not produce great quantities of contaminated waste. Most of all, it reduces the diagnostic time from at least 2 days for the bacterial isolation to less than 90 min. The assay is recommended as a rapid screening test of cholera cases caused by V. cholerae O139.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Faculty of Tropical Medicine, 420/6 Rajvithi Rd., Bangkok 10400, Thailand. Phone: 66-2-2460056, ext. 593. Fax: 66-2-2468340. E-mail: tmwcc{at}mahidol.ac.th.

Journal of Clinical Microbiology, December 1998, p. 3595-3600, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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