Direct immunofluorescence assay for rapid environmental detection of Vibrio cholerae O1
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
16475506
DOI
10.1007/bf02931428
Knihovny.cz E-resources
- MeSH
- Antigens, Bacterial immunology isolation & purification MeSH
- Escherichia coli immunology MeSH
- Fluorescent Antibody Technique, Direct methods MeSH
- Culture Media chemistry MeSH
- Water Microbiology * MeSH
- Bacterial Outer Membrane Proteins immunology isolation & purification MeSH
- Antibodies, Bacterial MeSH
- Salmonella typhi immunology MeSH
- Sensitivity and Specificity MeSH
- Shigella dysenteriae immunology MeSH
- Fresh Water microbiology MeSH
- Population Surveillance MeSH
- Vibrio cholerae O1 immunology isolation & purification MeSH
- Cross Reactions MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Antigens, Bacterial MeSH
- Culture Media MeSH
- Bacterial Outer Membrane Proteins MeSH
- Antibodies, Bacterial MeSH
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.
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