E-ISSN 2536-9520 | ISSN 1110-2047
 

Original Research (Original Article) 


Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat

Helmy A Torky, Hanaa A Ahmed, Emad S Omran.

Abstract
In this study, duplex real-time PCR approach with melting curve analysis is presented for the detection of Listeria monocytogenes, which are important food-borne bacterial pathogens usually present in imported frozen meat. Melting curve analysis using a SYBR Green I real-time PCR approach showed characteristic Tm values demonstrating the specific and efficient amplification of the two pathogens;73.2°C and 81.54°C for L. monocytogenes and Salmonella spp., respectively, the duplex, real-time PCR was equally sensitive to uniplex real-time PCR, using same amounts of purified DNA. When our established duplex real-time PCR assay was applied to artificially inoculated imported frozen meat the detection limit was 103 CFU/g for each of these pathogens without enrichment. The overall process included the culture enrichment based on the ISO standard, consisting of 24 h incubation in appropriate media (Half Fraser Broth for Listeria and buffered peptone water (BPW) for Salmonella), followed by a single DNA extraction of mixed enrichment aliquot. The results from this study showed that the developed duplex real-time PCR with melting curve analysis is promising as a rapid and cost-effective test method for improving food safety.

Key words: Salmonella, Listeria monocytogenes, Multiplex, Real-time PCR


 
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How to Cite this Article
Pubmed Style

Helmy A Torky, Hanaa A Ahmed, Emad S Omran. Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat. AJVS. 2015; 46(1): 100-109. doi:10.5455/ajvs.185240


Web Style

Helmy A Torky, Hanaa A Ahmed, Emad S Omran. Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat. http://www.alexjvs.com/?mno=185240 [Access: February 08, 2020]. doi:10.5455/ajvs.185240


AMA (American Medical Association) Style

Helmy A Torky, Hanaa A Ahmed, Emad S Omran. Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat. AJVS. 2015; 46(1): 100-109. doi:10.5455/ajvs.185240



Vancouver/ICMJE Style

Helmy A Torky, Hanaa A Ahmed, Emad S Omran. Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat. AJVS. (2015), [cited February 08, 2020]; 46(1): 100-109. doi:10.5455/ajvs.185240



Harvard Style

Helmy A Torky, Hanaa A Ahmed, Emad S Omran (2015) Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat. AJVS, 46 (1), 100-109. doi:10.5455/ajvs.185240



Turabian Style

Helmy A Torky, Hanaa A Ahmed, Emad S Omran. 2015. Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat. Alexandria Journal of Veterinary Sciences, 46 (1), 100-109. doi:10.5455/ajvs.185240



Chicago Style

Helmy A Torky, Hanaa A Ahmed, Emad S Omran. "Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat." Alexandria Journal of Veterinary Sciences 46 (2015), 100-109. doi:10.5455/ajvs.185240



MLA (The Modern Language Association) Style

Helmy A Torky, Hanaa A Ahmed, Emad S Omran. "Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat." Alexandria Journal of Veterinary Sciences 46.1 (2015), 100-109. Print. doi:10.5455/ajvs.185240



APA (American Psychological Association) Style

Helmy A Torky, Hanaa A Ahmed, Emad S Omran (2015) Duplex Real Time PCR for Simultaneous Detection of Salmonella Species and Listeria Monocytogenes in Frozen Meat. Alexandria Journal of Veterinary Sciences, 46 (1), 100-109. doi:10.5455/ajvs.185240





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