An immunofluorescence assay for direct detection of V. cholerae O1 was developed using polyclonal antibodies raised against outer membrane proteins (OMPs) of V. cholerae O1. Production of OMPs varied with growth media used; maximum production was found in tryptic soy broth. The detection system was specific because no cross-reactivity was observed with other bacteria including V. cholerae O139, E. coli, S. dysenteriae and Salmonella enterica subsp. enterica serovar Typhi. The technique was able to detect 240 CFU/mL of V. cholerae O1 suspended in phosphate-buffered saline. The assay coupled with bacterial enrichment in APW for 6 h detected as few as 5 CFU of V. cholerae in spiked samples. Moreover, a 2-h incubation of enriched bacterial cells in 0.1% yeast extract with 10 ppm nalidixic acid enhanced the bacterial size and helped in morphological identification of V. cholerae. Among 32 potable water samples from afflicted hand pumps and wells collected from a cholera-plagued area 12 were found to be contaminated with V. cholerae by immunofluorescence assay as well as by conventional culture methods. The proposed method could thus be employed in environmental surveillance of V. cholerae O1.

Biotechnology Division Defense Research and Development Establishment Gwalior 474 002 India

Zobrazit více v PubMed

Appl Environ Microbiol. 1987 Dec;53(12):2862-5 PubMed

J Bacteriol. 1959 Dec;78:886-91 PubMed

J Bacteriol. 1980 Oct;144(1):382-9 PubMed

Microbiol Mol Biol Rev. 1998 Dec;62(4):1301-14 PubMed

Microb Ecol. 1982 Dec;8(4):313-23 PubMed

J Bacteriol. 1981 Feb;145(2):1018-24 PubMed

J Clin Microbiol. 1994 Jan;32(1):249-52 PubMed

Clin Diagn Lab Immunol. 2001 Jul;8(4):768-71 PubMed

J Clin Microbiol. 1996 Dec;34(12):2968-72 PubMed

Clin Microbiol Rev. 1995 Jan;8(1):48-86 PubMed

Am J Epidemiol. 1982 Dec;116(6):959-70 PubMed

Lancet. 1993 Mar 13;341(8846):703-4 PubMed

Appl Environ Microbiol. 2002 Feb;68(2):995-8 PubMed

Appl Environ Microbiol. 1990 Aug;56(8):2370-3 PubMed

Microbiology (Reading). 1996 Aug;142 ( Pt 8):2181-6 PubMed

Appl Environ Microbiol. 1999 Mar;65(3):1348-51 PubMed

J Med Microbiol. 1995 Apr;42(4):251-7 PubMed

Infect Immun. 1992 Nov;60(11):4848-55 PubMed

Single multiplex polymerase chain reaction for environmental surveillance of toxigenic-pathogenic O1 and non-O1 Vibrio cholerae