Abstract
BackgroundIn late 2011, a new Orthobunyavirus of the Simbu serogroup named Schmallenberg virus (SBV) emerged in continental Europe. The virus is transmitted by hematophagous arthropods, with the Culicoides species as, so far known, main vectors. Infection with the virus can cause clinical signs in adult ruminants including diarrhea, fever and reduced milk production. Transplacental infection of the developing fetus can lead to malformations of varying severity. To assess seroprevalence of SBV in Sweden an indirect enzyme-linked immunosorbent assay (ELISA) was established in connection with the surveys. Here, we describe the development and evaluation of the indirect ELISA, based on whole virus as the coating antigen and a monoclonal antibody for the detection of antibodies to SBV in ruminant sera. The evaluation includes comparison between the in-house ELISA, virus neutralization test and an indirect commercial ELISA.ResultsThe optimal working dilutions of antigens and conjugate were estimated with checkerboard titrations. Comparative studies, including ROC analyses, were used for the selection of an optimal cut-off (S/P value = sample value as percentage of positive control value). With an estimated S/P value of 15% the whole virus ELISA showed a specificity of 100% and a sensitivity of 99.19% compared to virus neutralization test (VNT) and with a good consistency as shown in reproducibility and variability experiments. Furthermore, the comparison of our whole virus indirect ELISA to an indirect ELISA with a SBV nucleoprotein antigen, demonstrated a higher sensitivity of our test.ConclusionThe indirect whole virus ELISA described in this paper is a readily available test for serological analysis of SBV antibodies. Since this in-house ELISA demonstrates a specificity and sensitivity comparable to virus neutralization test and also shows a higher sensitivity compared to commercially available indirect ELISA, it is a useful alternative for surveillance and screening purposes of SBV.
Highlights
In late 2011, a new Orthobunyavirus of the Simbu serogroup named Schmallenberg virus (SBV) emerged in continental Europe
The present study describes the establishment and evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to SBV in cattle, sheep and goat sera
Both the pelleted cell debris fraction and the polyethylene glycol (PEG) concentrated virus were treated with Triton x-100 at three different concentrations of 0.1%, 0.5%, 1.0%, to explore the concentration required for an optimal solubilization and virus inactivation
Summary
In late 2011, a new Orthobunyavirus of the Simbu serogroup named Schmallenberg virus (SBV) emerged in continental Europe. Infection with the virus can cause clinical signs in adult ruminants including diarrhea, fever and reduced milk production. We describe the development and evaluation of the indirect ELISA, based on whole virus as the coating antigen and a monoclonal antibody for the detection of antibodies to SBV in ruminant sera. Since Schmallenberg virus (SBV) was identified in October 2011 [1] cases of SBV infection have been reported in ruminants in most European countries [2] and, as of late 2012, in Sweden [3]. The Schmallenberg virus is a member of the genus Orthobunyavirus within the family Bunyaviridae and related to the Simbu serogroup [1]. Results of full-genome and serologic investigations indicate that SBV belongs to the species Sathuperi virus and is not a reassortant but rather likely one of the ancestors of Shamonda virus [14]
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