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Laboratory Techniques Used to Maintain and Differentiate Biotypes of Vibrio cholerae Clinical and Environmental Isolates
Published on: May 30, 2017
Genetic characterization of Vibrio cholerae strains by inter simple sequence repeat-PCR
A Ravi Kumar1, V Sathish, G Balakrish Nair
1Laboratory of Molecular Genetics, Centre for DNA Fingerprinting and Diagnostics, Hyderabad, India.
FEMS Microbiology Letters
|May 25, 2007
Summary
Inter simple sequence repeat-PCR (ISSR-PCR) effectively characterizes Vibrio cholerae strains, distinguishing pathogenic from nonpathogenic types. This molecular tool aids in accurate phylogenetic classification and diagnostic genotyping of microbial pathogens.
Area of Science:
- Microbiology
- Genetics
- Molecular Biology
Background:
- Vibrio cholerae classification relies on serological methods.
- Distinguishing pathogenic from nonpathogenic strains is crucial for public health.
- Molecular techniques offer potential for enhanced phylogenetic analysis.
Purpose of the Study:
- To evaluate the utility of inter simple sequence repeat-PCR (ISSR-PCR) for characterizing Vibrio cholerae.
- To elucidate the phylogenetic relationships among pathogenic and nonpathogenic V. cholerae isolates.
- To compare ISSR-PCR with other molecular methods for V. cholerae genotyping.
Main Methods:
- Analysis of 45 V. cholerae strains (O1 El Tor, O139, non-O1/non-O139) using eight ISSR primers.
- Genomic DNA amplification targeting regions between simple sequence repeats (SSRs).
- Phylogenetic analysis using neighbor-joining method and confirmation of pathogenicity islands via PCR.
Main Results:
- ISSR-PCR clustered V. cholerae strains according to serogroup, with minor exceptions.
- Absence of pathogenicity islands confirmed the origin of outlier strains.
- ISSR-PCR provided finer resolution for toxinogenic subtypes compared to ERIC-PCR.
- Phylogenetic results were consistent with serological classification.
Conclusions:
- ISSR-PCR is an efficient method for phylogenetic classification of V. cholerae.
- The assay is valuable for diagnostic genotyping of microbial pathogens.
- ISSR-PCR offers a robust alternative for prokaryotic genome characterization.
