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[Identification of Vibrio cholerae O1 by flow cytometry]
F J Alvarado-Alemán1, C González-Bonilla, C Wong-Arambula
1Laboratorio de Citometría de Flujo, Instituto Nacional de Diagnóstico y Referencia Epidemiológicos, México, D.F., Mexico.
Revista Latinoamericana De Microbiologia
|October 1, 1994
Summary
Laser flow cytometry (LFC) rapidly detects Vibrio cholerae in water samples. This method shows high sensitivity and specificity, distinguishing V. cholerae O1 from other microorganisms within hours.
Area of Science:
- Microbiology
- Immunology
- Analytical Chemistry
Background:
- Vibrio cholerae poses a significant public health threat, necessitating rapid and accurate detection methods.
- Traditional culture methods for V. cholerae identification are time-consuming, delaying response to outbreaks.
- Advancements in cytometry offer potential for faster and more sensitive microbial detection.
Purpose of the Study:
- To evaluate the efficacy of laser flow cytometry (LFC) for the rapid detection of Vibrio cholerae in peptonated water samples.
- To compare the performance of polyclonal (PolAb) and monoclonal antibodies (MoAb) in LFC-based detection of V. cholerae.
- To assess the sensitivity and specificity of LFC for identifying V. cholerae O1 and Non O1 strains in complex environmental matrices.
Main Methods:
- Analysis of 72 peptonated water samples using laser flow cytometry (LFC).
- Direct fluorescence technique employing fluorescein-conjugated polyclonal (PolAb) and monoclonal antibodies (MoAb).
- Comparison of LFC results with positive culture methods for V. cholerae identification.
Main Results:
- Polyclonal antibodies detected 33 positive samples.
- Monoclonal antibodies demonstrated high specificity, with only three V. cholerae O1 false negatives out of 20 positive samples.
- All 13 V. cholerae Non O1 samples were accurately detected using MoAb.
- High correlation index (R=0.96) observed between LFC and pure V. cholerae O1 strains, compared to R=0.69 with autofluorescence of peptonated water samples.
Conclusions:
- Laser flow cytometry (LFC) offers a rapid and sensitive method for detecting Vibrio cholerae in environmental samples.
- Monoclonal antibodies provide high specificity for identifying V. cholerae O1 and Non O1 strains.
- LFC can distinguish V. cholerae O1 from mixed microbial populations in a few hours, outperforming traditional methods.