Development and evaluation of an immunochromatographic strip for the detection of serum antibodies against bluetongue virus.

Abstract

In this study, an immunochromatographic strip (ICS) was developed for the detection of bluetongue virus (BTV) serum antibodies. Colloidal gold particles labeled with streptococcal protein G (SPG), which can bind to the F(C) fragment of mammalian immunoglobulins, were used as the detector reagent. A recombinant VP7 BTV protein and a purified rabbit anti-SPG antibody were immobilized on test and control regions of a nitrocellulose membrane, respectively. In order to evaluate the ICS, 37 sera from animals exposed to different BTV serotypes were used as positive controls. In addition, 50 positive sera against viruses other than BTV, and eight sera taken from naive healthy sheep were used to determine the specificity of the ICS. Three hundred and three field sera taken from sheep and cattle were used after the above sera had been used for validation. Compared with the competitive ELISA (c-ELISA), the specificity and sensitivity of the ICS was 97.6% and 100%, respectively. There was excellent agreement between the results obtained by c-ELISA and the ICS (kappa=0.930). As it is rapid and easy to use, the test is suitable for the serological surveillance of BTV infection in the field.