A spectrophotometric method for evaluating a latex agglutination assay of Salmonella typhi lipopolysaccharide.
A spectrophotometer set at a wavelength of 400 nm was used to read reaction mixtures containing a 0.01% suspension of antibody-sensitized latex particles and different amounts of soluble antigen, after incubation in tubes for 30 min. The antigen used was Salmonella typhi lipopolysaccharide, and the antibody was an O-9-specific monoclonal antibody. Agglutination was indicated by a fall in the turbidimetric measurement compared to control, unagglutinated latex. It was observed that agglutination increased with increasing concentrations of antigen to a maximum at 0.25-1.0 micrograms/ml, after which, the amount of agglutination decreased at a similar rate as far as 1 mg/ml of antigen, when no agglutination occurred at all. Thus, a bi-symmetrical curve was obtained, suggestive of a precipitation reaction. At the 'equivalence point', the turbidity of the reaction mixture compared to control fell from OD400 1.1 to OD400 0.3. A similar inhibition of reaction at 'antigen excess' was observed visually in the reaction mixtures. Conventional slide tests performed in parallel could also be inhibited, but at higher antigen levels. Inhibition could be achieved with a commercially available latex used for the detection of Neisseria meningitidis antigen.
D000942:Antigens, Bacterial / Q000032:analysis*; D006801:Humans; D007841:Latex Fixation Tests / Q000295:instrumentation / Q000379:methods; D008070:Lipopolysaccharides / Q000032:analysis* / Q000652:urine; D009345:Neisseria meningitidis / Q000276:immunology; D012485:Salmonella typhi / Q000276:immunology*; D013053:Spectrophotometry / Q000379:methods
D000942:Antigens, Bacterial; D008070:Lipopolysaccharides