PCR-based identi¢cation ofVibrio cholerae and the closely related species Vibriomimicus using the large chromosomal ori sequence ofVibrio cholerae

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.

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    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 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.

    Item Type: Article
    URI: http://www.eprints.iicb.res.in/id/eprint/453
    Subjects: Infectious Diseases and Immunology
    Divisions: Indian Institute of Chemical Biology
    Depositing User: Mr Shyamal Nath
    Date Deposited: 01 Nov 2011 17:36
    Last Modified: 07 Feb 2012 11:31
    Official URL: htpp://dx.doi.org/10.1111/j.1574-6968.2006.00146.x
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