Multiplex PCR annealing temperature optimization for the detection of Listeria monocytogenes
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Abstract
Listeria monocytogenes is an opportunistic intracellular pathogen that has become one of the most important causes of foodborne infections worldwide. Due to the importance of L. monocytogenes in public health many protocols for the identification and typing of this organism have been described, but for the most part they are very complex and long lasting. For this reason, a multiplex PCR was developed for the rapid and sensitive detection of this bacterium in different types of samples. The objective of the present study was to optimize the multiplex PCR technique for the identification of L. monocytogenes. Different annealing temperatures were tested to determine which has the highest specificity, avoiding nonspecific amplifications. Amplifications were obtained at 57 ° C and lower temperatures. It is concluded that the best temperature for tm is 57 ° C.
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References
M. Doumith et al., «New aspects regarding evolution and virulence of Listeria monocytogenes revealed by comparative genomics and DNA arrays», Infect. Immun., vol. 72, n.o 2, pp. 1072–1083, 2004. https://iai.asm.org/content/72/2/1072.short
P. S. Mead et al., «Food-related illness and death in the United States.», Emerg. Infect. Dis., vol. 5, n.o 5, p. 607, 1999. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2627714/
C. M. de Noordhout et al., «The global burden of listeriosis: a systematic review and meta-analysis», Lancet Infect. Dis., vol. 14, n.o 11, pp. 1073–1082, 2014. https://www.sciencedirect.com/science/article/pii/S1473309914708709
R. L. Buchanan, L. G. Gorris, M. M. Hayman, T. C. Jackson, y R. C. Whiting, «A review of Listeria monocytogenes: an update on outbreaks, virulence, dose-response, ecology, and risk assessments», Food Control, vol. 75, pp. 1–13, 2017. https://www.sciencedirect.com/science/article/pii/S0956713516306892
G. Kooper, G. Calderón, S. Schneider, W. Domínguez, y G. Gutiérrez, Enfermedades transmitidas por alimentos y su impacto económico. Estudio de caso en Costa Rica, El Salvador, Guatemala, Honduras y Nicaragua. ORGANIZACIÓN DE LAS NACIONES UNIDAS PARA LA AGRICULTURA Y LA ALIMENTACIÓN. Roma. ORGANIZACIÓN DE LAS NACIONES UNIDAS PARA LA AGRICULTURA Y LA ALIMENTACIÓN Roma, 2009. https://www.sidalc.net/cgi-bin/wxis.exe/?IsisScript=MAGSV.xis&method=post&formato=2&cantidad=1&expresion=mfn=004727
J. D. Palumbo, M. K. Borucki, R. E. Mandrell, y L. Gorski, «Serotyping of Listeria monocytogenes by enzyme-linked immunosorbent assay and identification of mixed-serotype cultures by colony immunoblotting», J. Clin. Microbiol., vol. 41, n.o 2, pp. 564–571, 2003. https://jcm.asm.org/content/41/2/564.short
Y. Chen y S. J. Knabel, «Multiplex PCR for simultaneous detection of bacteria of the genus Listeria, Listeria monocytogenes, and major serotypes and epidemic clones of L. monocytogenes», Appl Env. Microbiol, vol. 73, n.o 19, pp. 6299–6304, 2007. https://aem.asm.org/content/73/19/6299.short
L. Lotfollahi, A. Chaharbalesh, M. A. Rezaee, y A. Hasani, «Prevalence, antimicrobial susceptibility and multiplex PCR-serotyping of Listeria monocytogenes isolated from humans, foods and livestock in Iran», Microb. Pathog., vol. 107, pp. 425–429, 2017. https://www.sciencedirect.com/science/article/pii/S0882401016305617
K. Núñez-Montero et al., «Listeria costaricensis sp. nov.», Int. J. Syst. Evol. Microbiol., 2018. DOI: https://doi.org/10.1099/ijsem.0.002596.
M. Doumith, C. Buchrieser, P. Glaser, C. Jacquet, y P. Martin, «Differentiation of the major Listeria monocytogenes serovars by multiplex PCR», J. Clin. Microbiol., vol. 42, n.o 8, pp. 3819–3822, 2004. https://jcm.asm.org/content/42/8/3819.short
A. Lievens, S. Jacchia, D. Kagkli, C. Savini, y M. Querci, «Measuring digital PCR quality: performance parameters and their optimization», PLoS One, vol. 11, n.o 5, p. e0153317, 2016. http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0153317