ABSTRACT
A rapid method for detection of the surface lipopolysaccharide antigen and the cells of the causative agent of bovine brucellosis was developed. The method represents a sandwich format immunochromatographic assay. The contact between the sample and the test strip with immobilized immunoreagents initiates the fluid movement along the membrane components of the test strip, immunochemical reactions, and the formation of colored bands. The novel method requires 10 minutes to determine the lipopolysaccharide antigen of the cell wall of the brucellosis causative agent at concentrations down to 10 ng/mL and the Brucella abortus cells at concentrations down to 10(6) cells/mL (5 x 10(4) cells in the sample). The specificity of the immunodetection was confirmed. The designed test system can be used for the rapid field diagnosis of brucellosis in cattle.
Subject(s)
Antibodies, Bacterial/immunology , Antigens, Bacterial/analysis , Brucella abortus/immunology , Brucellosis/diagnosis , Chromatography, Affinity , Lipopolysaccharides/analysis , Animals , Antibodies, Bacterial/chemistry , Antibodies, Immobilized/chemistry , Antibodies, Immobilized/immunology , Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/immunology , Brucella abortus/isolation & purification , Brucellosis/immunology , Brucellosis/microbiology , Cattle , Color , Mice , Reagent Kits, Diagnostic , Reagent Strips , Sensitivity and Specificity , Time FactorsABSTRACT
Two preparations based on monoclonal antibodies to bacteria of the genus Brucella have been obtained. From the monoclonal preparations and globulins isolated from them erythrocyte diagnostica have been obtained with the use of amidol. Experiments on the cross indication of brucellae and other bacteria by means of these diagnostica and a similar preparation obtained from polyclonal serum have shown very high specificity of erythrocyte immunoreagents prepared from monoclonal antibodies.
