Comparison of an LPS-specific competitive ELISA with a motility enrichment culture method (MSRV) for detection of Salmonella typhimurium and S. enteritidis in chickens.
We report here evaluation of a competitive enzyme-linked immunosorbent assay (c-ELISA) for detection of Salmonella spp. in chicken organs and faeces. The c-ELISA used a monoclonal antibody (MAb), specific for a genus-specific epitope of the outer core oligosaccharide of salmonellae. Salmonella lipopolysaccharide (LPS) in samples competed with Salmonella LPS coated on microtitre plates, for binding to the MAb. Competition reduced binding of the MAb to the LPS on the plate and of the secondary antibody to the MAb hence reducing the chromogenic signal. Stable coating and minimal false positive were achieved by conjugating LPS to poly-L-lysine. The c-ELISA was compared with motility enrichment culture using modified semisolid Rappaport Vassiliadis (MSRV) medium, which detected less than 10(2) CFU/g, and did not allow migration of non-salmonella species. The c-ELISA detected 10(6) CFU of enriched culture or 10(2)-10(3) CFU of Salmonella/g of faeces. Its limit of detection was thus higher than that of MSRV culture and it had a sensitivity of 92.9% and a specificity of 96.7%.
Tan
S
S
Department of Veterinary Microbiology and Immunology, University of Guelph, Ont, Canada. tans@em.agr.ca
Gyles
C L
CL
Wilkie
B N
BN
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001431:Bacteriological Techniques; D001667:Binding, Competitive; D002478:Cells, Cultured; D002645:Chickens; D003470:Culture Media; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D008070:Lipopolysaccharides / Q000378:metabolism; D009068:Movement; D011201:Poultry Diseases / Q000175:diagnosis* / Q000382:microbiology; D012481:Salmonella Infections, Animal / Q000175:diagnosis* / Q000382:microbiology; D012477:Salmonella enteritidis / Q000302:isolation & purification* / Q000502:physiology; D012486:Salmonella typhimurium / Q000302:isolation & purification* / Q000502:physiology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D003470:Culture Media; D008070:Lipopolysaccharides