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PCR-based identification of Vibrio cholerae and the closely related species Vibrio mimicus using the large chromosomal ori sequence of Vibrio cholerae
- Source :
- FEMS microbiology letters. 257(1)
- Publication Year :
- 2006
-
Abstract
- The bacterial chromosomal replication origin (ori) sequences are a highly conserved essential genetic element. In this study, the large chromosomal replication origin sequence of Vibrio cholerae (oriCIVC) has been targeted for identification of the organism, including the biotypes of serogroup O1. The oriCIVC sequence-based PCR assay specifically amplified an 890 bp fragment from all the V. cholerae strains examined. A point mutation in the oriCIVC sequence of the classical biotype of O1 serogroup led to the loss of a BglII site, which was utilized for differentiation from El Tor vibrios. Interestingly, the PCR assay amplified a similarly sized ori segment, designated as oriCIVM, from V. mimicus strains, but failed to produce any amplicon with other strains. Cloning and sequencing of the oriCIVM revealed high sequence similarity (96%) with oriCIVC. The results indicate that V. mimicus is indeed very closely related to V. cholerae. In addition, the BglII restriction fragment length polymorphism (RFLP) between oriCIVM and oriCIVC sequences allowed us to differentiate the two species. The ori sequence-based PCR-RFLP assay developed in this study appears to be a useful method for rapid identification and differentiation of V. cholerae and V. mimicus strains, as well as for the delineation of classical and El Tor biotypes of V. cholerae O1.
- Subjects :
- Sequence analysis
Molecular Sequence Data
Replication Origin
medicine.disease_cause
Microbiology
El Tor
Polymerase Chain Reaction
Vibrio mimicus
Species Specificity
Vibrionaceae
Genetics
medicine
Humans
Molecular Biology
Vibrio cholerae
biology
Base Sequence
Vibrio cholerae O1
Sequence Analysis, DNA
Amplicon
Chromosomes, Bacterial
biology.organism_classification
Vibrio
Bacterial Typing Techniques
Restriction fragment length polymorphism
Polymorphism, Restriction Fragment Length
Subjects
Details
- ISSN :
- 03781097
- Volume :
- 257
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- FEMS microbiology letters
- Accession number :
- edsair.doi.dedup.....488a1c455ccd5172fc73139df9928813