Development of a blocking-ELISA for the detection of antibodies against Histomonas meleagridis in chickens and turkeys.

Abstract

Histomonosis is an infectious disease of mainly galliform birds that can cause high mortality, especially in commercial turkey flocks. Several diagnostic tools were available to detect its causative agent, the flagellated protozoan Histomonas meleagridis. However, serological tools were largely lacking. Recently, an indirect ELISA has been described, but this test was not validated for specificity and might suffer from cross-reactivity with related protozoa. Therefore, a specific blocking-ELISA for the detection of antibodies against H. meleagridis in chicken and turkey sera was developed. For this purpose, monoclonal antibodies were raised against a detergent extracted protein of H. meleagridis. These MAbs bound to morphologically identified histomonads in liver tissue of an infected turkey. The MAbs were conjugated with horseradish peroxidase, after which the most reactive conjugate was used to set-up the blocking-ELISA. Experimentally infected turkeys (n=9) and chickens (n=10) seroconverted in the blocking-ELISA within 2-4 weeks following inoculation with a H. meleagridis field strain. The MAb did not bind to Tetratrichomonas gallinarum antigen and experimentally inoculated T. gallinarum seropositive layer chickens (n=18) showed the same inhibition percentages in the blocking-ELISA as negative control birds did. Therefore, it was concluded that H. meleagridis blocking-ELISA does not cross-react with T. gallinarum, which is a closely related protozoan frequently occurring in poultry. The repeatability and reproducibility of the H. meleagridis blocking-ELISA were high. The new blocking-ELISA is a promising tool for experimental studies; however, further validation of this test with field samples is necessary before it can be used for diagnostic purposes.