Detection of Salmonella typhimurium in Different Food Sources by Conventional Method and multiplex PCR Assay
DOI:
https://doi.org/10.53555/bp.v2i7.130Keywords:
Salmonella Spp, Salmonella typhimurium, Food, Beverage,, Culture method, mPCR.Abstract
The occurrence of foodborne diseases is increasing throughout the world. Bacteria of the genus Salmonella are responsible for food poisoning and, in some cases, may be fatal. The aim of this study was to adapt the multiplex PCR technique (mPCR) on the rapid and direct identification of the presence of Salmonella sp. and Typhimurium in different food sources. Salmonella detection by both conventional culture and mPCR methods were performed on 400 samples collected over a 7-month period between December 2013 and June 2014 from street vendors, exposed foods that are
sold on the sidewalks, and in popular restaurants. Salmonella isolates were identified by a multiplex-PCR using three sets of primers targeting the invA , Mdh and fliC genes sequences from Salmonella spp. and S. typhimurium respectively. The results of culture method indicated that 73 samples (18.25%) out of the 400 showed positive results, and displayed that (10)40% of the examined frozen meat, (9)36% of minced meat, (16) 64% of frozen chicken, (5)20% of hamburger, (6)24% of fresh kebab, (4)16% of salad and ice cream, (3)12% of each basturma, fruit Cocktail, orange juice and raisin juice, (2)8% of mayonnaise and tabbouleh were contaminated with Salmonella Spp., whilst pomegranate juice and watermelon were not contaminated. The traditional method for the detection of Salmonella reveals Salmonella and bacteria-like Salmonella, a Serological detection was used to distinguish the Salmonella only. The results indicate 61 samples (83.56 %) out of the 73 were Salmonella spp., and 13(30.14%) samples out of 61 were Salmonella typhimurium. The results of mPCR indicated that 61 samples (15.25%) out of the 400 demonstrated positive results for the invA target gene as Salmonella spp. The results displayed that (8)32% of the examined frozen meat, (13)52% of frozen chicken,
(6)24% of minced meat and fresh kebab, (4)16% of hamburger and salad, (3)12% of each basturma, Chickpea, fruit cocktail and raisin juice (2) 8% of each Mayonnaise, Tabbouleh, orange juice and ice cream were contaminated with Salmonella Spp., whilst pomegranate juice and watermelon not contaminated. On the other hand 22 samples (5.5%) out of the 400 demonstrated positive results for Mdh target gene as Salmonella typhimurium. The results displayed that(5) 20% of the examined frozen
meat, (7)28% of frozen chicken, (4)16% of minced meat (2)8% of each hamburger and fresh kebab, and (1)4% of basturma and salad were contaminated with Salmonella typhimurium, whilst other plant products, beverage and ice cream were not contaminated with Salmonella typhimurium. Multiplex PCR successfully amplified the DNA fragments corresponding in size as fallows Salmonella spp.389
bp (invA target gene), Salmonella typhimurium 261bp ( Mdh target gene ) , but not from all the non-Salmonella. fliC gene did not work when used with invA and Mdh genes, may be due to its large size and need different temperature cycles. Our conclusion was that the traditional method is laborious , time consuming and less accurate because it detects Salmonella and bacteria-like Salmonella. Whilst mPCR was found to be a very sensitive test that allowed rapid and reliable identification of
these bacteria in food and beverage samples. Results of this study can be used by agriculture and health organizations in Iraq.
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