PCR-based identi¢cation ofVibrio cholerae and the closely related species Vibriomimicus using the large chromosomal ori sequence ofVibrio cholerae
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PCR-based identi¢cation ofVibrio cholerae and the closely related species Vibriomimicus using the large chromosomal ori sequence
Mukhophadyay, Asish K
Bhadra, Rupak K
Infectious Diseases and Immunology
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 PCRRFLP 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.
Saha, Arjun and Deb, Reema and Shah, Sangita and Ramamurthy, Thandavarayan and Shinoda, Sumio and Mukhophadyay, Asish K and Bhadra, Rupak K (2006) PCR-based identi¢cation ofVibrio cholerae and the closely related species Vibriomimicus using the large chromosomal ori sequence ofVibrio cholerae. FEMS Microbiol Lett, 257 (1). pp. 84-91.