Development of a surface plasmon resonance-based assay for the detection of Corynebacterium pseudotuberculosis infection in sheep |
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Authors: | Sharon Stapleton Bernard Bradshaw |
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Affiliation: | a School of Biotechnology, Dublin City University, Dublin 9, Ireland b National Centre for Sensor Research, Dublin City University, Dublin 9, Ireland c Department of Agriculture and Food, Central Veterinary Research Laboratory, Backweston Campus, Young's Cross, Celbridge, Co. Kildare, Ireland |
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Abstract: | 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. |
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Keywords: | Corynebacterium pseudotuberculosis Caseous lymphadenitis Surface plasmon resonance (SPR) Serum Non-specific binding |
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