Development of a sample pooling strategy for the serodiagnosis of small ruminant lentiviral infections using the ELITEST-MVV ELISA

Abstract To reduce the costs of using the ELITEST-MVV, we explored the possibilities of sample pooling. Straight forward pooling applying the manufacturer's test conditions resulted in a significant loss of sensitivity. This was solved by using lower pre-dilutions of the samples than prescribed. Although an increase of background signal was encountered, discrimination between positive and negative samples was even better at pre-dilutions up to 12.5× as compared to the standard pre-dilution of 100×. This implied that pooling of up to eight samples was feasible. Receiver operating characteristic (ROC) analysis was used to determine the optimal cut-off value for the testing of pooled serum samples. A model for cost-benefit analysis of pooling was applied which combines the economics of the technical performance of the modified assay and other additional cost factors connected with pooling such as hands-on time for composing the pools, expected seroprevalence in the test population, sample tracing and testing the individual samples of positive pools. We concluded that pooling of samples was only feasible for monitoring SRLV-free accredited flocks because of their very low prevalence of infection. A pool consisting of five samples turned out to be the economical optimum although technically pool sizes of 10 samples were permitted.