Evaluation of serovar-independent ELISA antigens of Actinobacillus pleuropneumoniae in pigs, following experimental challenge with A. pleuropneumoniae, Mycoplasma hyopneumoniae and Pasteurella multocida

Objective To compare the sensitivity and specificity of six serological enzyme‐linked immunosorbent assays (ELISAs) based on serovar‐independent antigens of Actinobacillus pleuropneumoniae (App) and investigate cross‐reactivity in disease‐free pigs challenged with Mycoplasma hyopneumoniae and Pasteurella multocida. Design Five experimental pig trials using direct challenge with App serovars 1, 7 or 15 or direct challenge with M. hyopneumoniae and/or various dose rates of P. multocida. Procedure A 39‐kDa outer membrane protein antigen and five recombinant antigens from the apxIVA gene of App were evaluated. The latter were derived from the ApxIVA N‐terminus (ApxIVA‐N, ApxIVA‐NP, ApxIVA‐NPS) or C‐terminus (ApxIVA‐C, ApxIVA‐CP). Pigs were sampled after challenge and clinical and necropsy findings evaluated. Results The 39‐kDa ELISA had high sensitivity but lacked specificity, with significantly increased cross‐reactivity following P. multocida challenge. ELISAs based on ApxIVA N‐terminus antigens were significantly more sensitive than C‐terminus antigens for the detection of App‐induced disease. Although ApxIVA‐N and ApxIVA‐NP ELISAs had increased reactivity following P. multocida challenge, they retained high specificity for App‐induced disease (90–93%). Affinity purified ApxIVA‐NP antigen had marginally better specificity than ApxIVA‐N, without reduced sensitivity. Mycoplasma hyopneumoniae did not affect serological cross‐reactivity. In disease‐free pigs, the specificity of the ApxIVA‐NPS ELISA may be adversely affected by nasal carriage of apparently low‐virulence App strains. Conclusions ApxIVA‐N‐based ELISAs can be used for evaluating App status in commercial herds, but some appear limited by high carriage rates of low‐virulence App. The 39‐kDa antigen is only of merit in exclusion of App disease by negative serology.