Development and evaluation of an indirect enzyme-linked immunosorbent assay for serological detection of Schmallenberg virus antibodies in ruminants using whole virus antigen

Acta Veterinaria Scandinavica

Katarina Näslund²  Gunilla Blomqvist²  Caroline Vernersson²  Stéphan Zientara¹  Emmanuel Bréard¹  Jean F Valarcher² 

[1] Virology Unit, French Agency for Food Environmental and Occupational Health Safety, Maisons-Alfort, F-94703, France

[2] Department of Virology, Immunobiology and Parasitology, National Veterinary Institute, Uppsala, SE-75189, Sweden

关键词: Goat;    Sheep;    Cattle;    Antibody detection;    Schmallenberg virus;    Whole virus antigen;    Indirect ELISA;   

Others  :  1082521
DOI  :  10.1186/s13028-014-0071-1

 received in 2014-05-19, accepted in 2014-10-09,  发布年份 2014

【 摘 要 】

Background

In 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.

Results

The 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.

Conclusion

The 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.

【 授权许可】

   
2014 Näslund et al.; licensee BioMed Central Ltd.

【 图 表 】

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