Journal of Arak University of Medical Sciences

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Isolation of Listeria Monocytogenes from Meat Products and Study on PrfA Gene in Bacteria Isolated and Compaire Them with Clinical Samples and Standard Samples

Jamileh Nowroozi, Elham Siasi Torbati, Robab Baharvand,
Volume 16, Issue 9 (12-2013)

Abstract

Background: Listeria is a gram positive bacterium, facultive intracellular pathogene. Prf A gene has an important role in virulence. The purpose of this study was to determine of the presence of Listeria monocytogenes in various meat products and presence of prf A gene in bacteria isolated from food samples and compare them with clinical samples and standard samples.

Materials and Methods: During 6 months of 60 samples of beef, chicken, ham, cocktail, sausage, from a butchery’s Khoramabad and shops in Khoramabad and Tehran were collected. L. monocytogenes was isolated according to cold enrichment method and prf A gene were analyzed by PCR (polymerase chain reaction) method. Statistical analysis was performed with the software SPSS (statistical package for social science).

Results: From 60 samples of meat and meat products, 8 (13.3%) were positive for Listeria spp. Among in these samples, 4 cases of L. monocytogenes (6.6%), 3 cases (5%), L. innocua and 1 case (1.6%) L. welshimri, were isolated which L. innocua was isolated from meat and poultry samples and L. monocytogens from meat, chicken, ham and L. welshimeri from meat were isolated. Prf A gene was detected in isolated L. monocytogenes and donated bacteria from dairy products, clinical and standard samples, 2 cases of donated samples of vegetable contained prf A genes.

Conclusion: The prf A gene (activator of transcription and regulators the expression of other virulence genes), in the studied samples were observed. This gene plays a role in pathogenesis. Because these bacteria are transmitted through food and is a serious threat to public health. Thus identification of bacteria especially its genes could be possible to find some ways to prevent the bacteria and avoid disease from it.

Optimization of a Multiplex PCR for Simultaneous Detection of Foodborne Pathogens Salmonella spp. and Escherichia coli O157:H7 and Contamination Prevalence Assay in Meat Products

Hossein Morsali, Golnaz I Asaadi Tehran, Hanieh Asaadi, Sajjad Yazdansetad, Reza Najafpour,
Volume 20, Issue 6 (9-2017)

Abstract

Abstract
Background: Salmonella spp. and Escherichia coli O157:H7 are the most common bacterial foodborne pathogens contaminating food products especially meat. It is essential to detect the pathogens rapidly, specifically, and simultaneously by selection and optimization of suitable reference genes. The present study was conducted to simultaneously detect E. coli O157:H7 and Salmonella spp. in meat products and contamination prevalence assay by using multiplex PCR based on rfbE and invA genes amplification in Zanjan province, northwest of Iran.
Materials and Methods: A total of 74 meat samples were obtained from various regions of Zanjan province, randomly. 25 grams of each meat sample was completely homogenized in 225 ml of Mueller-Hinton broth growth medium and incubated. Bacterial strains were purified and DNA extraction was performed from purified bacterial isolates. Simultaneous amplification of rfbE and invA gene fragments was done with specific primers by optimization of a multiplex PCR. Finally, the sensitivity of the method was evaluated by inoculation of the bacteria to the meat.
Results: Out of 74 meat samples, 6(8%), 4(5.4%), and 2(2.7%) samples were positive for E. coli O157:H7, Salmonella, and both E. coli O157:H7-Salmonella, respectively. Multiplex PCR indicated high sensitivity for simultaneous detection of the pathogens in lowest dilution of the bacteria that had been inoculated to the meat.
Conclusion: In this study, a multiplex PCR was optimized based on Salmonella spp. and E. coli O157:H7 virulence genes for rapid and simultaneous detection of the pathogens with high sensitivity and specificity. Multiplex PCR as a reliable tool for rapid and simultaneous detection of foodborne pathogens to prevent contamination of food products.

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