Browse > Article

Detection of Salmonella in Milk by Sandwich ELISA using Anti-Outer Membrane Protein Immunoglobulins  

최석호 (상지대학교 생명공학과)
Publication Information
Food Science of Animal Resources / v.24, no.2, 2004 , pp. 176-181 More about this Journal
Abstract
The specificity of sandwich enzyme-linked immunosorbent assay (ELISA) to detect Salmonella in milk was determined in this study. The antibodies used in sandwich ELISA were egg yolk immunoglobulin G (IgY) obtained after immunization of hen with outer membrane protein (OMP) fraction from Salmonella typhimurium and rabbit IgG obtained after immunization of rabbit with the purified OMP with the molecular weight of 40,000. The immunoblot assay showed that the IgY reacted strongly with OMP with the molecular weight of 6,000 and the rabbit IgG reacted strongly with OMP with the molecular weights of 40,000, 35,000, and 6,000 from the bacteria including Salmonella which belongs to Enterobacteriaceae. The IgY and rabbit IgG also reacted with other proteins from Salmonella typhimurium in immunoblot assay. Competitive ELISA showed that IgY showed specifity to react with two strains of Salmonella typhimurium and Salmonella cholerasuis but not with Escherichia coli and Yersinia enterocolitica. Two strains of Salmonella typhimurium added to UHT milk showed the highest absorbance of all the bacteria used in the sandwich ELISA. Some strains of Salmonella cholerasuis showed higher absorbances than non-Salmonella bacteria.
Keywords
Salmonella; outer membrane protein; ELISA; milk;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Akita, E. M. and Nakai, S. (1992) Immunoglobulin from egg yolk: Isolation and purification. J. Food Sci. 57, 629-634   DOI
2 Isibasi, A., Oritz, V., Vargas, M., Pantagua, J., Gonzalez, C., Moreno, J., and Kumate, J. (1988) Protection against Salmonella typhi infection in mice after immunization with outer membrane proteins from Salmonella typhi 9. 12. d. Vi. Infect. Immun. 56, 2953-2959   PUBMED
3 Johnstone, A. and Thorpe, R. (1987) Immunochemistry in Practice 2nd ed. Blackwell Scientific Publications
4 Martinez-Flores, I., Cano, R., Bustamante, V. H., Calva, E., and Puente, J. L. (1999) The ompB operon partially determines differential expression of OmpC in Salmonella typhi and Escherichia coli. J. Bacteriol. 181, 556-562   PUBMED
5 McDonough, P. L., Jacobson, R. H., Timoney, J. F., Mutalib, A., Kradel, D. C., Chang, Y. F., Shin, S. J., Lein, D. H., Trock, S., and Wheeler, K. (1998) Interprtation of antibody responses to Salmonella enterica enteritidis gm flagellin in poultry flocks are enhanced by a kinetics-based enzyme-linked immunosorbent assay. Clin. Diag. Lab. Immunol. 5, 550-555
6 Sojka, M. G., Carter, M. A., and Thorns, C. J. (1998) Characterization of epitopes of type I fimbriae of salmonella using monoclonal antibodies of SEF21 fimbriae of Salmonella enteritidis. Vet. Microbiol. 59, 149-170
7 Arocklasamy, A. and Krishnaswamy, S. (2000) Purifica-tion of integral outer-membrane protein OmpC a surface antigen from Salmonella typhi for structure function studies: a method applicable to enterobacterial major outer- membrane protein. Anal. Biochem. 283, 64-70   DOI   ScienceOn
8 Liu, J.-L., Benson, C. E., Munro, D. S., and Wolf, B. (2001) Sensitive and rapid immunoassays for Salmonella enteritidis. J. Clin. Lab. Anal, 15, 165-170   DOI   ScienceOn
9 Udhayakumar, V. and Muthukkaruppan, V. R. (1987) Protective immunity induced by outer membrane proteins of Salmonella typhimurium in mice. Infect. Immun. 55, 1583-1587
10 Thorns, C. J., Bell, M. M., Sojka, M. G., and Nicholas, R. A. (1996) Development and application of enzyme-linked immunosorbent assay for specific detection of Salmonella enteritidis infections in chickens based on antibodies to SEF14 fimbrial antigen. J. Clin. Microbiol. 34, 792-797   PUBMED
11 Morisse, J. P. and Cotte, J. P. (1994) Evaluation of some risks factors in bovine salmonellosis. Vet. Res. 25, 185-191   PUBMED
12 Ng, S. P., Tsul, C. P., Roberts, D., Chen, P. Y., and Ng, M. H. (1996) Detection and serogroup differentiation of Salomonella spp. in food within 30 hours by enrichment-immunoassay with a T6 monoclonal antibody capture enzyme-linked immunosorbent assay. Appl. Environ. Microbiol. 62, 2294-2302   PUBMED
13 Wyatt, G. M., Lee, H. A., Dionysiou, S., Morgan, M. R. A., Stokely, D. J., AI-hajji, A. H., Richards, J., Sillis, A. J., and Jones, P. H. (1996) Comparison of a microtitration plate ELISA with a standard cultural procedure for the detection of Salmonella spp. in chicken. J. Food Prot. 59, 238-243   PUBMED
14 Yoshmatsu, M. A. and Zawistowski, J. (2001) Application of rapid dot blot immunoassy for detection of Salmonella enterica serovar enteritidis in eggs, poultry, and other foods. Appl. Environ. Microbiol. 67, 459-461   DOI   ScienceOn
15 Nikaido, H. (1994) Porins and specific diffusion channels in bacterial outer membranes. J. BioI. Chem. 269, 3905-3908
16 Phillips, J. D. and Griffiths, M. W. (1989) Pasteurized dairy products-the constraints imposed by environmental bacterial contamination. In Food Contamination from Environmental Sources, Advances in Environemntal Science and Technology. Nriagu, J. O. and Simmons, M. S.(eds.), pp. 387. John Wiley and Sons, NY
17 Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685   DOI   PUBMED   ScienceOn