Lateral flow assays for the detection of African swine fever virus antigen are not fit for field diagnosis of wild boar carcasses

Abstract

African swine fever (ASF) is one of the most important viral diseases of domestic pigs and wild boar. Apart from endemic cycles in Africa, ASF is now continuously spreading in Europe and Asia. As ASF leads to severe but unspecific clinical signs and high lethality, early pathogen detection is of utmost importance. Recently, 'point-of-care' (POC) tests, especially immunochromatographic assays, have been intensively discussed for the use in remote areas but also in the context of on-farm epidemiological investigations and wild boar carcass screening. The later topic was the starting point for our present study. In detail, we evaluated the performance of the commercially available INGEZIM ASFV CROM Ag lateral flow assay (Eurofins Technologies Ingenasa) with selected high-quality reference blood samples, and with blood samples from wild boar carcasses collected under field conditions in Germany. While we observed a sensitivity of roughly 77% in freeze-thawed matrices of close to ideal quality, our approach to simulate field conditions in direct testing of blood samples from carcasses without any modification, resulted in a drastically reduced sensitivity of only 12.5% with the given sample set. Freeze thawing increased the sensitivity to roughly 44% which mirrored the overall sensitivity of 49% in the total data set of wild boar carcass samples. A diagnostic specificity of 100% was observed. In summary, the antigen LFA should not be regarded as a substitute for any OIE listed diagnostic method and has very limited use for carcass testing at the point of care. For optimized LFA antigen tests, the sensitivity with field samples must be significantly increased. An improved sensitivity is seen with freeze-thawed samples, which may indicate problems in the accessibility of ASFV antigen that could be overcome, to a certain extent, with assay modifications.

Keywords: African swine fever virus; antigen lateral flow assay; point-of-care test; sensitivity; specificity.