Detection of Mannheimia haemolytica-Specific IgG, IgM and IgA in Sera and Their Relationship to Respiratory Disease in Cattle

Abstract

Mannheimia haemolytica is one of the major causes of bovine respiratory disease in cattle. The organism is the primary bacterium isolated from calves and young cattle affected with enzootic pneumonia. Novel indirect ELISAs were developed and evaluated to enable quantification of antibody responses to whole cell antigens using M. haemolytica A1 strain P1148. In this study, the ELISAs were initially developed using sera from both M. haemolytica-culture-free and clinically infected cattle, then the final prototypes were tested in the validation phase using a larger set of known-status M. haemolytica sera (n = 145) collected from feedlot cattle. The test showed good inter-assay and intra-assay repeatability. Diagnostic sensitivity and specificity were estimated at 91% and 87% for IgG at a cutoff of S/P ≥ 0.8. IgM diagnostic sensitivity and specificity were 91% and 81% at a cutoff of sample to positive (S/P) ratio ≥ 0.8. IgA diagnostic sensitivity was 89% whereas specificity was 78% at a cutoff of S/P ≥ 0.2. ELISA results of all isotypes were related to the diagnosis of respiratory disease and isolation of M. haemolytica (p-value < 0.05). These data suggest that M. haemolytica ELISAs can be adapted to the detection and quantification of antibody in serum specimens and support the use of these tests for the disease surveillance and disease prevention research in feedlot cattle.

Keywords: ELISA; IgA; IgG; IgM; Mannheimia haemolytica; antibody; bovine respiratory disease.

Figure 1

Distribution of M. haemolytica IgG, IgM, and IgA isotype-specific ELISA sample-to-positive (S/P; Y-axis) based on test validation samples (n = 145). Serum samples collected from the cattle that were negative for M. haemolytica culture on study day 0 (n = 100) were classified as true M. haemolytica antibody-negative. Serum samples collected from the cattle at least 14 days after M. haemolytica isolation positive (n = 45) were classified as true M. haemolytica antibody positive.

Figure 2

Receiver operating characteristic (ROC) curve analysis of the M. haemolytica IgG, IgM, and IgA isotype-specific ELISA sample-to-positive (S/P) based on testing serum samples from M. haemolytica-culture-free (true negative) (n = 100) and M. haemolytica-positive (true-positive) cattle (n = 45).