Open AccessPCR‐based identification of Vibrio cholerae and the closely related species Vibrio mimicus using the large chromosomal ori sequence of Vibrio cholerae
Author(s) -
Saha Arjun,
Deb Reema,
Shah Sangita,
Ramamurthy Thandavarayan,
Shinoda Sumio,
Mukhophadyay Asish K.,
Bhadra Rupak K.
Publication year - 2006
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2006.00146.x
Subject(s) - vibrio cholerae , biology , el tor , genetics , amplicon , restriction fragment length polymorphism , vibrionaceae , bglii , microbiology and biotechnology , sequence analysis , restriction enzyme , polymerase chain reaction , gene , hindiii , bacteria
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 ( oriCI VC ) has been targeted for identification of the organism, including the biotypes of serogroup O1. The oriCI VC sequence‐based PCR assay specifically amplified an 890 bp fragment from all the V. cholerae strains examined. A point mutation in the oriCI VC sequence of the classical biotype of O1 serogroup led to the loss of a Bgl II site, which was utilized for differentiation from El Tor vibrios. Interestingly, the PCR assay amplified a similarly sized ori segment, designated as oriCI VM , from V. mimicus strains, but failed to produce any amplicon with other strains. Cloning and sequencing of the oriCI VM revealed high sequence similarity (96%) with oriCI VC . The results indicate that V. mimicus is indeed very closely related to V. cholerae . In addition, the Bgl II restriction fragment length polymorphism (RFLP) between oriCI VM and oriCI VC 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.