ISSN 2736-1756
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 12 (6), pp. 001-005, June, 2018. © International Scholars Journals
Full Length Research Paper
Rapid, specific and concurrent detection of Listeria, Salmonella and Escherichia coli pathogens by multiplex PCR in Iranian food
Hamid Reza Tavakoli1, Ali Najafi2* and Ali Ahmadi2
1Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
2Research Center of Molecular Biology, Baqiyatallah University of Medical Sciences, Tehran, Iran
Accepted 05 March, 2018
Abstract
We are unable to detect all microorganisms in media. In consequence, molecular methods like PCR based techniques can mend our difficulties in this era. Herein, we describe simultaneous detection of major food-borne pathogens Listeria monocytogenes, Salmonella spp. and Escherichia coli O157:H7. Iranian food materials used for comparison of traditional microbiological methods (such as culture and serology) and multiplex PCR method in the detection of pathogens were prepared from several local restaurant, including eggs, raw milk, Raw kobide, salad, chicken, and cheese. Following DNA extraction, PCR assay were performed, using three specific primer pair. Because of all different sizes of the amplified fragments for each uniplex reaction, we optimized the each primers concentration to achieve a clearly visible band pattern of agarose gel (210 bp for Listeria, 556 bp for E. coli and 942 bp for Salmonella). In conclusion, uniplex and multiplex PCR was considered to perceive detection of the pathogens simultaneously.
Key words: Listeria monocytogenes, Salmonella spp., Escherichia coli O157H7, multiplex PCR.