Pooling serum to identify cohorts of nonmilking cattle likely to be infected with Bovine viral diarrhea virus by testing for specific antibodies

Testing for specific antibodies against Bovine viral diarrhea virus (BVDV) in pooled serum may present an opportunity to decrease the cost of screening for herds of high seroprevalence and increased likelihood of active infection. Experimental serum pools (n = 280) were created by combining equal aliquots of serum from between 5 and 25 individuals. A further 188 serum pools were generated from field serum samples. All pools and individual sera were tested for BVDV-specific antibodies by enzyme-linked immunosorbent assay (ELISA), according to manufacturer's instructions. Pools returned repeatable results, with coefficients of variation generally below 10%. The presence of serum from a persistently infected (PI) individual in the pool had no significant effect on the ELISA sample-to-positive (S/P) ratio. The results revealed that a single strong antibody-positive individual could maintain a positive result (at the manufacturer's threshold) in pools of up to 128, while even a single weak-positive animal would generate a positive result in pools of up to 8. The S/P ratio of the pool was positively related to the within-pool prevalence of antibody-positive individuals. However, as the strength of the individual positive animals contributing to the pool had a large effect on the pool S/P ratio, the S/P ratio could not be used to accurately predict the within-pool prevalence of field serum pools. An alternative method of S/P ratio interpretation was pursued, and a two-graph receiver operating characteristic analysis allowed segregation of pools into low, medium, and high risk with good results when applied to field serum pools.