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. 1996 Jul;62(7):2294–2302. doi: 10.1128/aem.62.7.2294-2302.1996

Detection and serogroup differentiation of Salmonella spp. in food within 30 hours by enrichment-immunoassay with a T6 monoclonal antibody capture enzyme-linked immunosorbent assay.

S P Ng 1, C O Tsui 1, D Roberts 1, P Y Chau 1, M H Ng 1
PMCID: PMC168010  PMID: 8779567

Abstract

We previously described an antigen capture enzyme-linked immunosorbent assay which makes use of monoclonal antibody T6, which recognizes an epitope on the outer core polysaccharide of Salmonella lipopolysaccharide molecules that is common to almost all Salmonella serovars. In this paper, we show that this assay can detect between 10(5) and 10(7) Salmonella cells per ml even in the presence of excess Escherichia coli. A total of 153 of 154 (99%) serogroup A to E strains and 51 of 78 (71%) serogroup F to 67 strains were reactive as determined by this assay. This corresponds to a detection rate of approximately 98% of all salmonellae known to affect humans. None of the 65 strains of non-Salmonella bacteria tested positive. Taking advantage of the O-factor polysaccharides also present on the antigen captured by the immobilized T6 antibody, we showed that 136 of 154 Salmonella serogroup A to E strains (88%) were correctly differentiated according to their serogroups by use of enzyme conjugates of a panel of O-factor-specific monoclonal antibodies. We evaluated this assay for the detection and serogroup differentiation of salmonellae directly from enrichment cultures of simulated food, eggs, pork, and infant formula milk. All 26 samples which had been contaminated with Salmonella spp. were detected by T6 (100% sensitivity), with only one false-positive result from 101 samples not contaminated by Salmonella spp. (99% specificity). The detection time was substantially reduced to between 17 and 29 h, depending on the enrichment methods used. Since there were no false-negative results, we concluded that this enrichment-immunoassay method can afford rapid screening for Salmonella spp. in food samples.

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Selected References

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  1. Avrameas S., Ternynck T. Peroxidase labelled antibody and Fab conjugates with enhanced intracellular penetration. Immunochemistry. 1971 Dec;8(12):1175–1179. doi: 10.1016/0019-2791(71)90395-8. [DOI] [PubMed] [Google Scholar]
  2. Blackburn C. W. Rapid and alternative methods for the detection of salmonellas in foods. J Appl Bacteriol. 1993 Sep;75(3):199–214. doi: 10.1111/j.1365-2672.1993.tb02767.x. [DOI] [PubMed] [Google Scholar]
  3. Choi D., Tsang R. S., Ng M. H. Sandwich capture ELISA by a murine monoclonal antibody against a genus-specific LPS epitope for the detection of different common serotypes of salmonellas. J Appl Bacteriol. 1992 Feb;72(2):134–138. doi: 10.1111/j.1365-2672.1992.tb01814.x. [DOI] [PubMed] [Google Scholar]
  4. Hoiseth S. K., Stocker B. A. Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature. 1981 May 21;291(5812):238–239. doi: 10.1038/291238a0. [DOI] [PubMed] [Google Scholar]
  5. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  6. Lee H. A., Wyatt G. M., Bramham S., Morgan M. R. Enzyme-linked immunosorbent assay for Salmonella typhimurium in food: feasibility of 1-day Salmonella detection. Appl Environ Microbiol. 1990 Jun;56(6):1541–1546. doi: 10.1128/aem.56.6.1541-1546.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lim P. L., Fok Y. P. Detection of group D salmonellae in blood culture broth and of soluble antigen by tube agglutination using an O-9 monoclonal antibody latex conjugate. J Clin Microbiol. 1987 Jul;25(7):1165–1168. doi: 10.1128/jcm.25.7.1165-1168.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lu G. Z., Tsang R. S., Chau P. Y., Choi D., Law D., Ng M. H. Characterization and application of a murine monoclonal antibody that reacts specifically with the serogroup D1 Salmonella. FEMS Microbiol Lett. 1991 May 15;64(2-3):135–140. doi: 10.1111/j.1574-6968.1991.tb04649.x. [DOI] [PubMed] [Google Scholar]
  9. Luk J. M., Chan K. H., Tsang R. S., Ng M. H. Characterisation and application of a murine monoclonal antibody specific for the serogroup C2 Salmonella. J Med Microbiol. 1988 Jun;26(2):115–119. doi: 10.1099/00222615-26-2-115. [DOI] [PubMed] [Google Scholar]
  10. Tsang R. S., Chan K. H., Chau P. Y., Wan K. C., Ng M. H., Schlecht S. A murine monoclonal antibody specific for the outer core oligosaccharide of Salmonella lipopolysaccharide. Infect Immun. 1987 Jan;55(1):211–216. doi: 10.1128/iai.55.1.211-216.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Tsang R. S., Chan K. H., Lau N. W., Choi D. K., Law D. K., Ng M. H. Characterization of murine monoclonal antibodies against serogroup B salmonellae and application as serotyping reagents. J Clin Microbiol. 1991 Sep;29(9):1899–1903. doi: 10.1128/jcm.29.9.1899-1903.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Tsang R. S., Chan K. H., Lau N. W., Ng M. H. Production and characterization of murine monoclonal antibodies specific for serogroups E1 and E4 Salmonella. Diagn Microbiol Infect Dis. 1990 Nov-Dec;13(6):453–460. doi: 10.1016/0732-8893(90)90076-8. [DOI] [PubMed] [Google Scholar]
  13. Tsang R. S., Nielsen K., Henning M. D., Schlecht S., Aleksić S. A murine monoclonal antibody that recognizes a genus-specific epitope in the Salmonella lipopolysaccharide outer core. Zentralbl Bakteriol. 1991 Jan;274(4):446–455. doi: 10.1016/s0934-8840(11)80080-0. [DOI] [PubMed] [Google Scholar]
  14. Wong S. S., Yuen K. Y., Yam W. C., Lee T. Y., Chau P. Y. Changing epidemiology of human salmonellosis in Hong Kong, 1982-93. Epidemiol Infect. 1994 Dec;113(3):425–434. doi: 10.1017/s0950268800068436. [DOI] [PMC free article] [PubMed] [Google Scholar]

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