Evaluation of a multiplex immunoassay for bovine respiratory syncytial virus and bovine coronavirus antibodies in bulk tank milk against two indirect ELISAs using latent class analysis

Bovine respiratory syncytial virus (BRSV) and bovine coronavirus (BCV) are responsible for respiratory disease and diarrhea in cattle worldwide. The Norwegian control program against these infections is based on herd-level diagnosis using a new multiplex immunoassay. The objective of this study was to estimate sensitivity and specificity across different cut-off values for the MVD-Enferplex BCV/BRSV multiplex, by comparing them to a commercially available ELISA, the SVANOVIR® BCV-Ab and SVANOVIR® BRSV-Ab, respectively.

We analyzed bulk tank milk samples from 360 herds in a low- and 360 herds in a high-prevalence area. As none of the tests were considered perfect, estimation of test characteristics was performed using Bayesian latent class models. At the manufacturers’ recommended cut-off values, the median sensitivity for the BRSV multiplex and the BRSV ELISA was 94.4 [89.8–98.7 95% Posterior Credibility Interval (PCI)] and 99.8 [98.7–100 95% PCI], respectively.

The median specificity for the BRSV multiplex was 90.6 [85.5–94.4 95% PCI], but only 57.4 [50.5–64.4 95% PCI] for the BRSV ELISA. However, increasing the cut-off of the BRSV ELISA increased specificity without compromising sensitivity. For the BCV multiplex we found that by using only one of the three antigens included in the test, the specificity increased, without concurrent loss in sensitivity.

At the recommended cut-off this resulted in a sensitivity of 99.9 [99.3–100 95% PCI] and specificity of 93.7 [88.8–97.8 95% PCI] for the multiplex and a sensitivity of 99.5 [98.1–100 95% PCI] and a specificity of 99.6 [97.6–100 95% PCI] for the BCV ELISA.