DETECCIÓN DE ESCHERICHIA COLI O157:H7 EN CARNE FRESCA DE RES MEDIANTE PCR MÚLTIPLEX
Abstract
Para la detección de E. coli O157:H7 en carne fresca de res, se comparó el método microbiológico tradicional y dos ensayos de PCR Múltiplex. En ensayos con muestras inoculadas se obtuvo una diferencia significativa entre PCR Múltiplex y aislamiento microbiológico. PCR Múltiplex mostró porcentaje mayor de sensibilidad, precisión relativa e índice kappa en el enriquecimiento con el caldo Ecm+n en comparación al CST+ccv. En 40 muestras de res corte tipo americano se detectaron 2 positivas (5%) por PCR múltiplex, mientras que por el método microbiológico no se logró su aislamiento en ninguna muestra.
Abstract
In the present study were compared the traditional microbiological method and two tests of multiplex PCR for the detection of E. coli O157:H7 in fresh beef meat. A significant difference was obtained between PCR multiplex and microbiological isolation in inoculated samples; in addition, in multiplex PCR was obtained a higher percentage of sensitivity, precision relative and Kappa index using the Ecm+n broth in comparison to the CST+ccv in the enrichment. In 40 samples of meat cuts American type were detected two positive samples (5%) by Multiplex PCR, whereas by the microbiological method was not obtained the isolation in any sample.
Palabras clave: PCR Múltiplex, E. coli O157:H7, Carne, Beef
Downloads
References
Blanco, M., J. E. Blanco, J. Blanco, E. A. González and M. P. Alonso. 1996. Prevalence and characteristics of human and bovine verotoxigenic Escherichia coli strains isolated in Galicia (north-western Spain). Eur. J. Epidemiol.; 12:13-19.
Arias-Echandi, M.L. 1996. Escherichia coli O157:H7. Laboratorio de Alimentos y Aguas de la Facultad de Microbiología. Universidad de Costa Rica.
USDA-FSIS, 1996. Revision 4 of Laboratory Communication 38 Protocol for Isolation and Identification ofEscherichia coli O157:H7. Amelia K. Sharar and Bonnie E. Rose.
Witham P.K., C. T. Yamashiro, K. J. Livak and C. A. Batt. 1996. A PCR-based assay for the detection of Escherichia coli shiga-like toxin genes in ground beef. Appl. Environ.Microbiol. 62:1347-1353.
Venkateswaran, K., Y. Kamijoh, E. Ohashi and H. Nakanishi. 1997. A simple filtration technique to detect Enterohemorrhagic Escherichia coli O157:H7 and its toxins in beef by multiplex PCR. Appl. Environ. Microbiol. 63:4127-4131.
Blanco, M., et. al., Op.cit.
Gannon, V. P. J., M. Rashed., R. K. King., and E. J. Golsteyn 1993. Detection and characterization of the eae gene of shiga-like toxin-producing Escherichia coli using polymerase chain reaction. J. of Clin. Microbiol. 31: 1268-1274.
Desmarchelier, P. M., S. S. Bilge., N. Fegan., L. Mills., J. C. Vary. Jr., and P. I. Tarr. 1998. A PCR specific forEscherichia coli O157:H7 based on the rfb locus encoding O157 lipopolisaccharide. J. Clin. Microbiol. 36: 1801-1804.
Gannon, V. P. J., S. D’souza., T. Graham., R. K. King., K. Rahn., and S. Read. 1997. Use of the flagellar H7 gene as a target in multiplex PCR assays and improved specificity in identification of enterohemorrhagic Escherichia coliStrains. J. Clin. Microbiol. 35: 656–662.
Doyle, J.J., and Doyle, J.L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19, 11-15.
Ferrán-Aranaz, M. 1996. SPSS para Windows. Programación y Análisis Estadístico. Serie McGraw-Hill de Informática. McGraw-Hill / Interamericana de España, S. A. ISBN: 84-481-0589-3.. pp: 580.
Malorny, B., J. Hoorfar, C. Bunge and R. Helmuth. 2003. Multicenter validation of the analytical accuracy ofSalmonella PCR: towards an international standard. Appl. Environ. Microbiol. 69(1):290-296.
Wang, G., C. G. Clark and F. G. Rodgers. 2002. Detection in Escherichia coli of the genes encoding the major virulence factors, the genes defining the O15:H7 serotype and components of the type 2 shiga toxin family by multiplex PCR. J. Clin. Microbiol. 40:3613-19.
Cebula, T. A., W. L. Payne and P. Feng. 1995. Simultaneous identification of strains of Escherichia coli serotype O157:H7 and their shiga-like toxin type by mismatch amplification mutation assay-multiplex PCR. J. Clin. Microbiol. 33:248-250.
Weagant, S. D. and K. G. Jinneman. 1995. An improved rapid technique for isolation of Escherichia coli 0157:H7 from foods. J. Food Prot. 58:7-12.
Campbell, G.R., J. Prosser, A. Glover and K. Killham. 2001. Detection of Escherichia coli O157:H7 in soli and water using multiplex PCR. J. Appl. Microbiol. 91:1004-1010.
Kehl, S. C. 2002. Minireview: Role of the laboratory in the diagnosis of enterohemorrhagic Escherichia coliinfections. J. Clin. Microbiol. 40(8): 2711-2715.
Fratamico P. M., S. K. Sackitey, M. Wiedmann and M. Yi Deng. 1995. Detection of Escherichia coli O157:H7 by Multiplex PCR. J. Clin. Microbiol. 33:2188-2191.
Jothikumar, N. and M. W. Griffiths. 2002. Rapid detection of Escherichia coli O157:H7 with multiplex realtime PCR assays. Appl. Environ. Microbiol. 68(6):3169-3171.
Ramotar, K., B. Waldhart, D. Church, R. Szumski and T. J. Louie. 1995. Direct detection of verotoxinproducingEscherichia coli in stool samples by PCR. J. Clin. Microbiol. 33(3):519-524.
Willshaw, G. A., H. R. Smith, D. Roberts, J. Thiriwell, T. Cheasty and B. Rowe.1993. Examination of raw beef products for the presence of Vero cytotoxin producing Escherichia coli, particularly those of serogroup O157. J. Appl. Bacteriol. 75:420-426.
Paton, A. W. and J. Paton. 1999. Direct detection of shiga toxigenic Escherichia coli strains belonging to serogroups O111, O157 and O113 by multiplex PCR. J. Clin. Microbiol. 37(10):3362-3365.
Downloads
Published
How to Cite
Issue
Section
License
The rights of the work belong to the author or authors, however, by sending it for publication in the Public Health and Nutrition Magazine of the Faculty of Public Health and Nutrition of the Autonomous University of Nuevo León, they grant the right for its first publication in between electronic, and possibly, in print to the Public Health and Nutrition Magazine. The license used is the Creative Commons attribution, which allows third parties to use what is published whenever the authorship of the work is mentioned and the first publication that is in the Public Health and Nutrition Magazine. Likewise, the author or authors will take into account that it will not be allowed to send the publication to any other journal, regardless of the format. The authors will be able to make other independent and additional contractual agreements for the non-exclusive distribution of the version of the article published in the Public Health and Nutrition Magazine (e.g., institutional repository or publication in a book) provided they clearly state that The work was published for the first time in the Public Health Magazine, Magazine of the Faculty of Public Health and Nutrition of the Autonomous University of Nuevo León.
