Molecular identification and characterization ofVibriospp. in different water samples of Lucknow region, Uttar Pradesh using Multiplex PCR

Abstract

The present study focused on the incidence of Vibrio spp. in water bodies used for human consumption and industrial purpose. As Vibrios constituted of both pathogenic and non-pathogenic species, a rapid method, the multiplex PCR assay was used to differentiate between them. The water collected from the 15 sampling sites was analyzed for their physico-chemical properties. Isolation of bacteria was carried out on selective media (TCBS) where both green and yellow colonies were observed. These colonies were cultured separately on non selective media. All the pure cultures obtained were used for the extraction of DNA using phenol - chloroform method, visualized on agarose gel and quantified using double beam spectrophotometer. Samples with higher yield of DNA were used for amplification using species specific primers by multiplex PCR. The results were documented and the study revealed the presence of non-pathogenic vibrio species in most of the water bodies, while pathogenic forms were present in few of them. However, the percentage of the pathogenic species was found to be more than non-pathogenic species. The percentage of pathogenic and non-pathogenic bacteria was 64.2% and 28.5% and 7.3% of both types were found in equal measures in the samples.