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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2020)
[Effective from January 1, 2020]
For more details click here

ICV 2018: 95.39
Index Copernicus ICI Journals Master List 2018 - IJCMAS--ICV 2018: 95.39
For more details click here

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Original Research Articles

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

Int.J.Curr.Microbiol.App.Sci.2020.9(2): 1532-1540
DOI: https://doi.org/10.20546/ijcmas.2020.902.176


Multiplex PCR for Detection of Escherichia coli O157 : H7
in Faeces from Sheep and Goat
Pralhad*, Basavaraj Awati, Arun Kharate, Pradeep Kumar, Srikanth Kulkarni,C. Mallinath, B. K. Ramesh and P. Suryakanth

Department of Veterinary Microbiology, Veterinary College, Bidar-585401, India


*Corresponding author
Abstract:

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.


Keywords: coli O157:H7; Food borne pathogen; Virulent Genes; Multiplex PCR; Sheep and Goat
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How to cite this article:

Pralhad, Basavaraj Awati, Arun Kharate, Pradeep Kumar, Srikanth Kulkarni , K. C. Mallinath, B. K. Ramesh and Suryakanth, P. 2020 Multiplex PCR for Detection of Escherichia coli O157 : H7 in Faeces from Sheep and Goat. Int.J.Curr.Microbiol.App.Sci. 9(2): 1532-1540. doi: https://doi.org/10.20546/ijcmas.2020.902.176