A pH sensitive, loop-mediated isothermal amplification assay for detection of Salmonella in food
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Abstract
Salmonella is a common foodborne pathogen. This study aimed to develop a rapid detection method for Salmonella spp. in food by using a pH sensitive, loop-mediated isothermal amplification (LAMP) assay. Firstly, a suitable pH indicator was selected to follow the change in pH of LAMP reactions. Then, the best primer set was selected from among three previously approved primer sets specific to the genes invA, invE, and phoP. The amplification-positive reactions were easily detectable with neutral red at a concentration of 15 mM (color change from faint orange to red). The best primers were those specific to phoP when incubated at 63.5°C for 1.5 h. The developed assay was validated for detection of Salmonella Typhimurium in spiked minced chicken meat and liquid egg with a sensitivity of 2 CFU/g or 2 CFU/mL. With this novel, pH-sensitive LAMP assay, Salmonella can be detected in food in 6 h.
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Almeida, C., Cerqueira, L., Azevedo, N., and Vieira, M. (2013). Detection of Salmonella enterica serovar Enteritidis using real time PCR, immunocapture assay, PNA FISH and standard culture methods in different types of food samples. International Journal of Food Microbiology, 161(1), 16-22.
Barbau-Piednoir, E., Bertrand, S., Mahillon, J., Roosens, N. H., and Bottledoorn, N. (2013). SYBR® green qPCR Salmonella detection system allowing discrimination at the genus, species and subspecies levels. Applied Microbiology and Biotechnology, 97(22), 9811-9824.
Bird, P., Flannery, J., Crowley, E., Agin, J., Goins, D., and Jechorek, R. (2014). Evaluation of the 3M™ Petrifilm™ Salmonella express system for the detection of Salmonella species in selected foods: collaborative study. Journal of AOAC International, 97(6), 1563-1575.
Chen, S., Yee, A., Griffiths, M., Larkin, C., Yamashiro, C. T., and Behari, R. (1997). The evaluation of a fluorogenic polymerase chain reaction assay for the detection of Salmonella species in food commodities. International Journal of Food Microbiology, 35(3), 239-250.
Chen, Z., Zhang, K., Yin, H., Li, Q., Wang, L., and Liu, Z. (2015). Detection of Salmonella and several common Salmonella serotypes in food by loop-mediated isothermal amplification method. Food Science and Human Wellness, 4(2), 75-79.
Eriksson, E., and Aspan, A. (2007). Comparison of culture, ELISA and PCR techniques for Salmonella detection in faecal samples for cattle, pig and poultry. BMC Veterinary Research, 3(21), 1-19.
Food and Drug Administration (FDA). (2017). BAM Chapter 5: Salmonella. U.S. Food and Drug Administration, [Online URL: https://www.fda.gov/food/foodscienceresearch/laboratorymethods/ucm070149.htm] accessed on May 20, 2017.
Goto, M., Honda, E., Ogura, A., Notomo, A., and Hanaki, K. (2009). Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue. Biotechniques. 46(3), 167-172.
Isogai, E., Makungu, C., Yabe, J., Sinkala, P., Nambota, A., and Isogai, H. (2005). Detection of Salmonella invA by isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN) in Zambia. Comparative Immunology, Microbiology and Infectious Diseases, 28(5-6), 363-370.
Jacobsen, C. S., and Holben, W. E. (2007). Quantification of mRNA in Salmonella sp. seeded soil and chicken manure using magnetic capture hybridization RT-PCR. Journal of Medical Microbiology, 69(2), 315-321.
Jothikumar, P., Narayanan, J., and Hill, V. R. (2014). Visual endpoint detection of Escherichia coli O157:H7 using isothermal genome exponential amplification reaction (GEAR) assay and malachite green. Journal of Microbiological Methods, 98, 122-127.
Kim, Y., and Lee, J. L. (2016). Rapid detection of Salmonella enterica serovar Enteritidis from eggs and chicken meat by real-time recombinase polymerase amplification in comparison with the two-step real-time PCR. Journal of Food Safety, 36(3), 402-411.
Kirk, M. D., Pires, S. M., Black, R. E., Caipo, M., Crump, J. A., Devleesschauwer, B., and Angulo, F. J. (2015). World Health Organization estimates of the global and regionald burden of 22 foodborne bacterial, protozoal, and viral diseases, 2010: a data synthesis. PLoS Medicine, 12(12), e1001921.
Kothary, M. H., and Babu, U. S. (2001). Infective dose of foodborne pathogens in volunteers: a review. Journal of Food Safety, 21(1), 49-73.
Kumar, P. P., Agarwal, R. K., Avinash, R. D., Prasannavadhana, A., Thomas, P., Kumar, A., Kataria, J. L., and Kerketta, P. (2014). Loop mediated isothermal amplification (LAMP) test for rapid detection of Salmonella in the chicken meat. Journal of Veterinary Public Health, 12(1), 7-12.
Li, X., Zhang, S., Zhang, H., Zhang, L., Tao, H., Yu, J., Zheng, W, Liu, C., Lü, D., Xiang, R., and Liu, Y. (2009). A loop-mediated isothermal amplification method targets the phoP gene for the detection of Salmonella in food samples. International Journal of Food Microbiology, 133(3), 252-258.
Malorny, B., Bunge, C., and Helmuth, R. (2007). A real-time PCR for the detection of Salmonella Enteritidis in poultry meat and consumption eggs. Journal of Medical Microbiology, 70(2), 245-251.
Nowak, B., von Müffling, T., Chaunchom, S., and Hartung, J., (2007). Salmonella contamination in pigs at slaughter and on the farm: a field study using an antibody ELISA test and a PCR technique. International Journal of Food Microbiology, 115(3), 259-267.
Nucera, D. M., Maddox, C. W., Hoien-Dalen, P., and Weigel, R. M. (2006). Comparison of API 20E and invA PCR for identification of Salmonella enterica isolates from swine production units. Journal of Clinical Microbiology, 44(9), 3388-3390.
Ohtsuka, K., Yanagawa, K., Takatori, K., and Hara-Kudo, Y. (2005). Detection of Salmonella enterica in naturally contaminated liquid eggs by loop-mediated isothermal amplification, and characterization of Salmonella isolates. Applied and Environmental Microbiology, 71(11), 6730-6735.
Popoff, M. Y., Bockemühl, J., and Brenner, F. W. (2000). Supplement 1998 (no. 42) to the Kauffmann-White scheme. Research in Microbiology, 151(1), 63-65.
Tanner, N. A., Zhang, Y., and Evans Jr., T. C. (2015). Visual detection of isothermal nucleic acid amplification using pH-sensitive dyes. Biotechniques, 58(2), 59-68.
Techathuvanan, C., and D'Souza, D. H. (2012). Reverse-transcriptase loop-mediated isothermal amplification as a rapid screening/monitoring tool for Salmonella enterica detection in liquid whole eggs. Journal of Food Science, 77(4), M200-M205.
Techathuvanan, C., Draughon, F. A., and D'Souza, D. H. (2010). Loop-mediated isothermal amplification (LAMP) for the rapid and sensitive detection of Salmonella typhimurium from pork. Journal of Food Science, 75(3), M165-M172.
Tomita, N., Mori, Y., Kanda, H., and Notomi, T. (2008). Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nature Protocols, 3(5), 877-882.
Wang, B., Huang, X., Ma, M., Shi, Q., Z., and Cai, Z. (2014). A simple quantum dot-based fluoroimmunoassay method for selective capturing and rapid detection of Salmonella Enteritidis on eggs. Food Control, 35(1), 26-32.
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(1), 112.
Ye, Y., Wang, B., Huang, F., and Song Y. S. (2011). Application of in situ loop-mediated isothermal amplification method for detection of Salmonella in foods. Food Control, 22(3-4), 438-444.
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(18), 6495-6501.