International Journal of Pure & Applied Bioscience (IJPAB)
Year : 2016, Volume : 4, Issue : 5
First page : (114) Last page : (120)
Article doi: http://dx.doi.org/10.18782/2320-7051.2371
Development of Duplex PCR for the Identification of Shigella and Salmonella: Major Water Borne Bacteria
Arunava Das*, M. Suruthi, N. Sri Nivetha and A. Reshma
Molecular Diagnostics and Bacterial Pathogenomics Research Laboratory, Department of Biotechnology, Bannari Amman Institute Technology, Sathyamangalam-638401, Erode District, Tamil Nadu, India
*Corresponding Author E-mail: arunavadas@bitsathy.ac.in
Received: 11.09.2016 | Revised: 23.09.2016 | Accepted: 25.09.2016
Abstract
The present study was focused to develop a duplex PCR for the identification of two major pathogens Shigella and Salmonella from water. A total of 12 waste water samples were collected from sewage treatment plant located in Erode district, Tamilnadu. These water samples were investigated for both microbiological and molecular studies. Standard biochemical test were able to identify tentatively 8 (66.6%) and 11(91.66%) isolates of Shigella and Salmonella respectively. The 23.1Kb genomic DNA were isolated from the bacteria and the protocol was standardized. Duplex polymerase chain reaction was then carried out to identify the presence of the Shigella invasive gene (invC; 875) and Salmonella enterotoxin gene (stn; 617bp). In dPCR out of 12 samples invC gene was detected from 6 (50%) samples andstn gene was detected from 7(58.3%) samples. Virulence gene specific dPCR developed in this study was very sensitive and accurate for the identification of pathogenic strains of Shigella and Salmonella from waste water. In SEM study, the ultra-structure of both the pathogens was visualized. As an outcome of the current study, the virulence genes like invCand stncan be used as a gene marker in PCR assays for the rapid diagnosis of pathogenic strains of Shigella and Salmonella respectively. Present study suggested that PCR is a sensitive, reliable tool over the traditional biochemical tests that are laborious and also time consuming. The study examines the risk of humans exposed to these zoonotic organisms via contaminated water from the sewage treatment plants and may result in serious water-borne illness in humans.
Key words: Salmonella, Shigella, sewage water, PCR, virulence.
Full Text : PDF; Journal doi : http://dx.doi.org/10.18782
Cite this article: Das, A., Suruthi, M., Nivetha, N.S. and Reshma, A., Development of Duplex PCR for the Identification of Shigella and Salmonella: Major Water Borne Bacteria, India , Int. J. Pure App. Biosci.4(5): 114-120 (2016). doi: http://dx.doi.org/10.18782/2320-7051.2371
