Genetic characterization ofVibrio choleraestrains by inter simple sequence repeat-PCR

Abstract

The utility of inter simple sequence repeat-PCR (ISSR-PCR) assay in the characterization and elucidation of the phylogenetic relationship between the pathogenic and nonpathogenic isolates of Vibrio cholerae is demonstrated. A total of 45 V. cholerae strains including 15 O1 El Tor, nine O139 and 21 non-O1/non-O139 strains were analyzed using eight ISSR primers. These primers, which are essentially simple sequence repeats (SSR) with additional nonrepeat bases at the 5' or 3' end, amplify genomic regions interspersed between closely spaced SSRs. Neighbor-joining analysis showed that the strains belonging to the same serogroup clustered together with the exception of one O1 and two O139 strains. The absence of pathogenicity islands in these strains, as confirmed by PCR, suggested their non-O1/non-O139 origin. Thus the ISSR-PCR-based phylogeny was consistent with the classification of V. cholerae based on serological methods. A finer resolution of the clustering of the toxinogenic O1 El Tor and toxinogenic O139 subtypes was obtained by ISSR-PCR analysis as compared with the Enterobacterial Repetitive Intergenic Consensus sequences-based PCR analysis for the same set of strains. Thus, it is proposed that ISSR-PCR is an efficient tool in phylogenetic classification of prokaryotic genomes in general and diagnostic genotyping of microbial pathogens in particular.

Green synthesis of silver nanoparticles derived from lemon and pomegranate peel extracts to combat multidrug-resistant bacterial isolates

“…Also, the polymorphism information content (PIC) was calculated as 0.231 generated from the SDS-PAGE pattern. Therefore, our results was similar with [ 40 ] reporting that the genetic similarity and protein profile of the isolated field German strains were compared to Mycoplasma bovis reference strain using SDS-PAGE. In addition, [ 39 ] examined the total proteins of 20 isolates that were taken from the NIH Islamabad using SDS-PAGE to determine the variability in the gene pool of E. coli , as shown in Fig.…”

Molecular identification and genetic diversity among Photorhabdus and Xenorhabdus isolates

“…Successful use of PCR-based SSR amplification followed by amplicon size determination to analyze the spread of microbial pathogens has been reported for Haemophilus influenzae and Candida albicans (Bretagne et al 1997; van Belkum et al 1997). Furthermore, the utility of inter-simple sequence repeat-PCR (ISSR-PCR) assay in the characterization and elucidation of the phylogenetic relationship between the pathogenic and nonpathogenic isolates of Vibrio cholerae was demonstrated (Kumar et al 2007). They proposed that ISSR-PCR is an efficient tool in phylogenetic classification of prokaryotic genomes in general and diagnostic genotyping of microbial pathogens in particular.…”