Enliven: Immunology and Immunotechniques

A Multispecies Serological Microarray to Detect Exposure of Wildlife to Multiple Pathogens
Author(s): Cawthraw S1*, Ruettger A2, Fenner J1, Ehricht R3,10, Muller E3,10, Gavier-Widen D4,5, Uhlhorn H4, Mawhinney I1, Birch C1, Gortazar C11, Tang Y1, Hutchings M6, Hannant D7, Billinis C8, Fimme Jan van der Wal9, Sachse K2, Petrovska L1

Wildlife is a major reservoir for many human and livestock pathogens. Wildlife sampling is often very opportunistic and restricted. Many commercial immunoassays have not been licensed for routine diagnostic use in wildlife and standardized measurement of the antibody levels to different antigens has not been performed. As part of the EU-funded Wild Tech project and with the aim of providing the basis for a diagnostic test for use in surveillance studies, we evaluated the use of miniaturized protein A/G multiplex ELISA-based microarrayfor the detection of specific antibodies against multiple pathogens in single serum samples from wild mammal species.Sera obtained from wild cervids and wild boar were tested by array and conventional serodiagnostic assays. Using samples identified as positive or negative by conventional methods, we determined diagnostic cut-off points for cervids and wild boar sera tested by array for specific antibodies against 34 different bacterial and viral antigens. Of these 34, tests for Mycobacterium bovis (TB), Brucella, Leptospira, Toxoplasma, Trichinella, Yersinia, Campylobacter and Salmonella antigens produced comparable results with the standard serological tests. As a measure of agreement between array values and standard tests, Cohen kappa values for antigens tested with deer sera ranged from 0.41 to 0.92 and for wild boar sera from 0.43 to 1 (where 0 means no agreement and 1 means complete agreement). Despite the lack of success with some antigens, the promising results with others demonstrate proof-of-principle that this approach to rapid serodiagnosis could finally yield tests of great value for wildlife disease surveillance.