National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
Department of Veterinary Microbiology, Veterinary College, Bidar-585401, India
coli O157:H7 is an emerging food borne pathogen having zoonotic significance. Though the primary reservoir of this serotype is cattle; Sheep and goat are also considered to be the main reservoir for E. coli O157:H7. Conventional cultural methods are time consuming method and also the specificity is very less. Multiplex PCR (mPCR) tends to be specific, more rapid and reliable. In the present study fecal samples collected from sheep (n=517) and goat (n=450) in different farms across Kalyana Karnataka region were analysed. The samples were processed and analysed for the cultural isolation, biochemical Characterization and latex agglutination test. An mPCR method for the detection and molecular Characterization of E. coli O157:H7 was standardized using primer pairs targeting six specific virulent genes; fliCh7, eaeA, rfbE, hly, stx1, and stx2. The mPCR produced species-specific amplicons of the six targeted genes of the size; 625 bp, 397bp, 296 bp, 166 bp, 210 bp and 484 bp, respectively. The percent of sheep and goats that showed shedding of E. coli O157 in the faeces was 3.86% (20 out of 517) and 2.88% (13 out of 450) respectively. The results obtained show that the standardized mPCR protocol is a rapid, highly sensitive, species-specific and reliable method for the detection of the pathogenic E. coli O157:H7 and could be used for identification and molecular Characterization of E. coli O157:H7 in suspected food and water borne outbreaks, disease investigations and routine analysis etc.
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