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Background   Klebsiella pneumonia , a Gram-negative bacterium belonging to the genus  Enterobacter , causes many human and livestock diseases. Notably, infected goats may develop pneumonia, septicemia, which can lead to occasional death, resulting in great economic losses in goat-farming industry. However, there are little systematic methods for detection of goat  Klebsiella pneumoniae  in livestock production.    Results  In this study, we developed a  Klebsiella pneumoniae  goat polyclonal antibody and established an indirect ELISA method to detect the  Klebsiella pneumoniae . After screening and optimizing the conditions for detection, we determined the optimal working dilutions of the coated-bacterial antigen, the polyclonal antibody, and the enzyme-labeled secondary antibody that were 1:800 (2.99 × 10 7  CFU/ml), 1:6400, and 1:5000, respectively. The optimal condition of coating and blocking were both 4 °C for 12 h. The optimal dilution buffers of bacterial antigen, the antibodies, and the blocking buffer were 0.05 mol/L carbonate buffer, 1% BSA phosphate buffer, and 1.5% BSA carbonate buffer, respectively. The cut-off value was determined to be 0.28, and the analytical sensitivity was 1:800 (dilution of a positive sample). Furthermore, there was no cross-reaction between the coated antigen and goat serum positive for antibodies against other bacteria, indicating that indirect ELISA could detect  Klebsiella pneumoniae  specifically in most cases. The average coefficients of variation of intra-assay and inter-assay were 4.37 and 5.17% indicating favorable reproducibility of indirect ELISA. In the detection of clinical veterinary samples, the positive rate of indirect ELISA was 6.74%, higher than that of conventional agglutination assays.    Conclusions  Taken together, we successfully established an indirect ELISA method for detecting antibodies against  Klebsiella pneumoniae  in goats, which can be applied in production.

Establishment and evaluation of an indirect ELISA for detection of antibodies to goat Klebsiella pneumonia