International Journal of Food Science and Agriculture

ISSN Print: 2578-3467 Downloads: 122162 Total View: 2265717
Frequency: quarterly ISSN Online: 2578-3475 CODEN: IJFSJ3

Evaluation of Loop-Mediated Isothermal Amplification for the Detection of Salmonella from Poultry Matrices

Raj Rajagopal1,*, Gabriela Lopez-Velasco1, John M. David1, Melissa Sisemore2, Jamie Goseland2

13M Food Safety, 3M Company, St. Paul, MN 55144, USA.

2WBA Analytical Laboratories, Inc., Springdale, AR 72762, USA.

*Corresponding author: Raj Rajagopal

Published: December 10,2021


Loop-mediated isothermal amplification (LAMP) has emerged as an alternative to PCR based methods for detection of food-borne pathogens, offering simple, easy to use, detection technology with high speed, efficiency, sensitivity and specificity. The performance of LAMP-bioluminescent assay as an alternative method for the detection of Salmonella in primary production samples, poultry rinses and raw poultry products compared to PCR was evaluated. After enrichment, boot swabs from poultry farms, carcass rinses and raw poultry products were tested by a LAMP-bioluminescent and a PCR assay. The LAMP-bioluminescent assay was able to detect Salmonella in the various matrices tested and had higher or equivalent sensitivity and specificity to the PCR method used. No significant difference (95% confidence interval) was found between the LAMP and PCR method as determined by probability of detection analysis. The Salmonella LAMP-bioluminescent assay enabled reliable and rapid detection of Salmonella in variety of poultry matrices and is an acceptable alternative to the PCR method.


[1] CDC.  (2021).  Salmonella and food., Accessed October 25, 2021. 

[2] Scallan, E., Hoekstra, R. M., Angulo, F. J., Tauxe, R. V., Widdowson, M. A., Roy, S. L., Jones, J. L., and Griffin, P. M. (2011).  Foodborne illness acquired in the United States—major pathogens.  Emerging infectious diseases, 17: 7-15. 

[3] Scharff, R. L. (2012).  Economic burden from health losses due to foodborne illness in the United States.  Journal of Food Protection, 75: 123-31. 

[4] Russell, S. (2012).  Controlling Salmonella in poultry production and processing, CRC press, Boca Raton, Florida. 

[5] WHO.  (2015).  WHO estimates of the global burden of foodborne diseases: foodborne disease burden epidemiology reference group 2007-2015., Accessed date: October 25, 2021. 

[6] Rouger, A., Tresse, O., and Zagorec, M. (2017).  Bacterial contaminants of poultry meat: sources, species, and dynamics.  Microorganisms, 5: 50. 

[7] USDA FSIS.  (2015).  Changes to the Salmonella and Campylobacter verification testing program: proposed performance standards for Salmonella and Campylobacter in not-ready to-eat comminuted chicken and turkey products and raw chicken parts and related agency verification procedures and other changes to agency sampling.  FSIS-2014-0023, Federal Register, 80: 3940-3950. 

[8] USDA FSIS.  (2019).  Changes to the Campylobacter verification testing program: revised performance standards for Campylo-bacter in not-ready-to-eat comminuted chicken and turkey and related agency procedures.  FSIS-2018-0044, Federal Register, 84: 38203-38210. 

[9] USDA FSIS.  (2021).  USDA launches new effort to reduce Salmonella illnesses linked to poultry.  Press release no.  0223.21., Accessed October 30, 2021. 

[10] Jasson, V., Jacxsens, L., Luning, P., Rajkovic, A., and Uyttendaele, M. (2010).  Alternative microbial methods: an overview and selection criteria.  Food Microbiology, 27: 710-730. 

[11] Mangal, M., Bansal, S., Sharma, S. K., and Gupta, R. K. (2016).  Molecular detection of foodborne pathogens: a rapid and accurate answer to food safety.  Critical Reviews in Food Science and Nutrition, 56: 1568-1584. 

[12] Park, S. H., Aydin, M., Khatiwara, A., Dolan, M. C., Gilmore, D. F., Bouldin, J. L., Ahn, S., and Ricke, S. C. (2014).  Current and emerging technologies for rapid detection and characterization of Salmonella in poultry and poultry products.  Food Microbiology, 38: 250-262. 

[13] Souii, A., M´hadheb-Gharbi, M. B., and Gharbi, J. (2016).  Nucleic acid-based biotechnologies for food-borne pathogen detection using routine time-intensive culture-based methods and fast molecular diagnostics.  Food Science and Biotechnology, 25: 11-20. 

[14] Wiedmann, M., Wang, S., Post, L., and Nightingale, K. (2014).  Assessment criteria and approaches for rapid detection methods to be used in the food industry.  Journal of Food Protection, 77: 670-690. 

[15] Mori, Y. and Notomi, T. (2009).  Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases.  Journal of Infection and Chemotherapy, 15: 62-69. 

[16] Mori, Y., Kanda, H., and Notomi, T. (2013).  Loop-mediated isothermal amplification (LAMP): recent progress in research and development.  Journal of Infection and Chemotherapy, 19: 404-411. 

[17] Notomi, T., Mori, Y., Tomita, N., and Kanda, H. (2015).  Loop-mediated isothermal amplification (LAMP): principle, features, and future prospects.  Journal of Microbiology, 53: 1-5. 

[18] Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N., and Hase, T. (2000).  Loop-mediated isothermal amplification of DNA.  Nucleic Acids Res., 28: E63. 

[19] Kaneko, H., Kawana, T., Fukushima, E., and Suzutani, T. (2007).  Tolerance of loop-mediated isothermal amplification to a culture medium and biological substances.  Journal of Biochemistry and Biophysical Methods, 70: 499-501. 

[20] Niessen, L., Luo, J., Denschlag, C., and Vogel, R. F. (2013).  The application of loop-mediated isothermal amplification (LAMP) in food testing for bacterial pathogens and fungal contaminants.  Food Microbiology, 36: 191-206.

[21] Plutzer, J. and Karanis, P. (2009). Rapid identification of Giardia duodenalis by loop-mediated isothermal amplification (LAMP) from faecal and environmental samples and comparative findings by PCR and real-time PCR methods. Parasitology Research, 104: 1527-1533. 

[22] Yang, Q., Wang, F., Prinyawiwatkul, W., and Ge, B. (2014). Robustness of Salmonella loop-mediated isothermal amplification assays for food applications. Journal of Applied Microbiology, 116: 81-88.

[23] Yang, Q., Domesle, K. J., and Ge, B. (2018). Loop-mediated isothermal amplification for Salmonella detection in food and feed: current applications and future directions. Foodborne Pathogens and Disease, 15: 309-331.

[24] Domesle, K. J., Yang, Q., Hammack, T. S., and Ge, B. (2018). Validation of a Salmonella loop-mediated isothermal amplification assay in animal food. International Journal of Food Microbiology, 264: 63-76.

[25] Gandelman, O. A., Church, V. L., Moore, C. A. Kiddle, G., Carne, C. A., Parmar, S., Jalal, H., Tisi, L. C., and Murray, J. A. (2010). Novel bioluminescent quantitative detection of nucleic acid amplification in real-time. PLoS ONE, 5: e14155.

[26] Bird, P., Flannery, J., Crowley, E., Agin, J. R., and Monteroso, L. (2016). Evaluation of the 3M™ Molecular Detection Assay (MDA) 2 - Salmonella for the detection of Salmonella spp. in select foods and environmental surfaces: collaborative study, first action 2016.01. Journal of AOAC International, 99: 980-997.

[27] Hu, L., Ma, L. M., Zheng, S., He, X., Wang, H., Brown, E. W., Hammack, T. S., and Zhang, G. (2017). Evaluation of 3M Mo-lecular Detection System and ANSR Pathogen Detection System for rapid detection of Salmonella from egg products. Poultry Science, 96: 1410-1418.

[28] Hu, L., Deng, X., Brown, E. W., Hammack, T. S., Ma, L. M., and Zhang, G. (2018). Evaluation of Roka Atlas Salmonella method for the detection of Salmonella in egg products in comparison with culture method, real-time PCR and isothermal amplification assays. Food Control, 94: 123-131.

[29] Huo, J., Huang, Y., and Rajagopal, R. (2021). Loop-mediated isothermal amplification vs. Guobiao standards method for detection of Salmonella in yoghurt and yoghurt-based drinks. International Journal of Dairy Science, 16: 90-97.

[30] Lim, H. S., Zheng, Q., Miks-Krajnik, M., Turner, M., and Yuk, H. G. (2015). Evaluation of commercial kit based on loop-mediated isothermal amplification for rapid detection of low levels of uninjured and injured Salmonella on duck meat, bean sprouts, and fishballs in Singapore. Journal of Food Protection, 78: 1203-1207.

[31] Rajagopal, R., Barnes, C. A., David, J. M., Goseland, J., and Goseland, J. (2021). Evaluation of a commercial loop-mediated isothermal amplification assay, 3MTM Molecular Detection Assay 2 - Campylobacter, for the detection of Campylobacter from poultry matrices. British Poultry Science, 62: 404-413.

[32] Sarowska, J., Frej-Mądrzak, M., Jama-Kmiecik, A., Kilian, A., Teryks-Wołyniec, D., and Choroszy-Król, I. (2016). Detection of Salmonella in foods using a reference PN-ISO method and an alternative method based on loop-mediated isothermal amplification coupled with bioluminescence. Advances in Clinical and Experimental Medicine, 25: 945-950.

[33] Yang, Q., Domesle, K. J., Wang, F., and Ge, B. (2016). Rapid detection of Salmonella in food and feed by coupling loop-mediated isothermal amplification with bioluminescent assay in real-time. BMC Microbiology, 16: 112.

[34] NPIP. (2019). National Poultry Improvement Plan Program Standards, USDA Animal and Plant Health Inspection Service Veterinary Services., Accessed October 25, 2021. 

[35] USDA FSIS MLG 4.11. (2021). Isolation and identification of Salmonella from meat, poultry, pasteurized egg, and siluriformes (fish) products and carcass and environmental sponges. 2021-08/MLG-4.11.pdf, Accessed October 25, 2021.

[36] Eijkelkamp, J. M., Aarts, H. J. M., and van der Fels-Klerx, H. J. (2009). Suitability of rapid detection methods for Salmonella in poultry slaughterhouses. Food Analytical Methods, 2: 1-13.

[37] Wehling, P., LaBudde, R. A., and Nelson, M. T. (2011). Probability of Detection (POD) as a statistical model for the validation of qualitative methods. Journal of AOAC International, 94: 335-347.

[38] Hu, L., Ma, L. M., Zheng, S., He, X., Hammack, T. S., Brown, E. W., and Zhang, G. (2018). Development of a novel loop-mediated isothermal amplification (LAMP) assay for the detection of Salmonella ser. Enteritidis from egg products. Food Control, 88: 190-197.

[39] Zhang, G., Brown, E. W., and González-Escalona, N. (2011). Comparison of real-time PCR, reverse transcriptase real-time PCR, loop-mediated isothermal amplification, and the FDA conventional microbiological method for the detection of Salmonella spp. in produce. Applied and Environmental Microbiology, 77: 6495-6501.

[40] Forstner, M. (2016). Salmonella flip book. Minnesota Department of Agriculture. FoodScienceResearch/RFE/UCM517352.pdf, Accessed date: October 25, 2021. 

[41] USDA FSIS MLG 8.13. (2021). Isolation and identification of Listeria monocytogenes from red meat, poultry, ready-to-eat siluriformes (fish) and egg products, and environmental samples. 2021-09/MLG-8.13.pdf, Accessed October 25, 2021.

How to cite this paper

Evaluation of Loop-Mediated Isothermal Amplification for the Detection of Salmonella from Poultry Matrices

How to cite this paper: Raj Rajagopal, Gabriela Lopez-Velasco, John M. David, Melissa Sisemore, Jamie Goseland. (2021) Evaluation of Loop-Mediated Isothermal Amplification for the Detection of Salmonella from Poultry MatricesInternational Journal of Food Science and Agriculture5(4), 717-727.