Lipopolysaccharide O-antigen antibody-based detection of the fish pathogen Flavobacterium psychrophilum.
Several yellow-pigmented species within the family Flavobacteriaceae are commonly associated with diseases in fish and are difficult to speciate due to their fastidious, slow-growing nature and cross-reactive antigens. Here we report the development of specific, antibody-diagnostic tests for Flavobacterium psychrophilum, the aetiological agent of rainbow trout fry syndrome and bacterial cold water disease. A unique antigen from F. psychrophilum, the lipopolysaccharide (LPS) O-polysaccharide (O-PS), formed the basis for the antibody test. LPS O-PS was purified and conjugated to keyhole limpet haemocyanin and bovine serum albumin for the generation of rabbit immune sera and the development of antibody-based diagnostic tests. Rabbit polyclonal anti-O-PS serum was highly specific for F. psychrophilum, without the need for prior cross-absorption with related bacteria and was the basis of an effective ELISA diagnostic test. Antibodies were purified from rabbit anti-O-PS serum and adsorbed onto coloured latex beads for the development of a specific, bead agglutination assay for F. psychrophilum.
Crump
Elizabeth M
EM
Department of Biochemistry and Microbiology, University of Victoria, Victoria, Canada.
Perry
Malcolm B
MB
Gale
Stephen
S
Crawford
Evan
E
Kay
William W
WW
D000907:Antibodies, Bacterial / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005417:Flavobacterium / Q000276:immunology* / Q000302:isolation & purification*; D007106:Immune Sera; D007841:Latex Fixation Tests; D008969:Molecular Sequence Data; D019081:O Antigens / Q000737:chemistry / Q000276:immunology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D007106:Immune Sera; D019081:O Antigens