Development of a surface plasmon resonance-based assay for the detection of Corynebacterium pseudotuberculosis infection in sheep.
Caseous lymphadenitis (CLA), a disease affecting sheep and goats, is caused by Corynebacterium pseudotuberculosis and is difficult to detect, especially at early stages in its development. A surface plasmon resonance-based biosensor assay for the detection of antibodies to the phospholipase D (PLD) exotoxin of C. pseudotuberculosis in sheep serum was successfully generated. It employed a recombinant form of PLD, which was immobilised, and all aspects of the assay including minimisation of non-specific binding, and the regeneration of the chip, were optimised. The applicability of the assay was initially demonstrated using sera collected from experimentally infected sheep and from sheep with no prior history of infection. The assay was then evaluated on a panel of clinical samples and the results obtained compared very favourably to those obtained by a double sandwich ELISA (over 90% similarity) and clearly verified its analytical value.
Stapleton
Sharon
S
School of Biotechnology, Dublin City University, Dublin 9, Ireland.
Bradshaw
Bernard
B
O'Kennedy
Richard
R
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D015374:Biosensing Techniques / Q000379:methods*; D003354:Corynebacterium Infections / Q000175:diagnosis / Q000662:veterinary*; D016925:Corynebacterium pseudotuberculosis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006041:Goats; D010739:Phospholipase D / Q000276:immunology / Q000378:metabolism; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis*; D020349:Surface Plasmon Resonance / Q000379:methods*
D000907:Antibodies, Bacterial; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins; D010739:Phospholipase D