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Background   Haemophilus parasuis  is a commensal pathogen in the swine upper respiratory tract and causes Glässer’s disease. Surveillance, screening for infection, and vaccination response of  H. parasuis  is hindered by the lack of a rapid antibody detection method.    Results  In the present study, a monomeric autotransporter was identified as a novel antigen for developing an indirect ELISA. The autotransporter passenger domain (Apd) was expressed, purified, and demonstrated to be specific in ELISA and western blotting. Mouse antiserum of recombinant Apd (rApd) recognized native Apd in the 15 serotype reference strains and five non-typeable isolate stains, but showed no reaction with seven other bacterial pathogens. The rApd ELISA was optimized and validated using 67 serum samples with known background, including 27 positive sera from experimentally infected and vaccinated pigs along with 40 negative sera that had been screened with  H. parasuis  whole cell ELISA from clinically healthy herds. The rApd ELISA provided positive and negative percent agreements of 96.4 and 94.9%, respectively, and an AUC value of 0.961, indicating that the assay produced accurate results.    Conclusion  Apd was a universal antigen component among 15 serotype and non-typeable strains of  H. parasuis  and was also specific to this pathogen. The rApd ELISA could detect antibodies elicited by  H. parasuis  infection and vaccination, thereby exhibiting the potential to be applied for Glässer’s disease diagnosis,  H. parasuis  vaccination evaluation, and large-scale serological surveillance.

Development and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domain