Monoclonal antibodies to bovine coronavirus and their use in enzymoimmunoanalysis and immunochromatography

Abstract

Two monoclonal antibodies (MAb) to the outer structural protein E2 (spike peplomeric protein) and two MAb to the inner capsid protein N of bovine coronavirus (BCV) were prepared and identified by Western blotting to be used for increasing the specificity and sensitivity of BCV detection. The MAb were checked by the haemagglutination inhibition test and immunoperoxidase tests and no cross reactivity with rotavirus was demonstrated by the immunoperoxidase test and ELISA. A mixture of all the four MAb at predetermined optimum concentrations was first used in sandwich ELISA and then, in combination with an anti‑coronavirus polyclonal antibody, for the development of a simple and rapid immunochromatographic test (ICT). The results of which can be read visually within 10 min. The inclusion of MAb into ELISA and ICT allows the detection of both intact and incomplete BCV virions. ELISA and ICT were used in the examination of a set of 74 faecal samples collected from calves suffering from diarrhoea. ELISA, used as the golden standard verified by electron microscopy, detected BCV in 15 samples (20.3%) and ICT in 16 samples. Three of the ICT‑positive samples were negative by ELISA. On the other hand, two of the 58 ICT‑negative samples were positive by ELISA. Sensitivity and specificity of ICT were 94.9% and 86.7%, respectively

Keywords

• immunochromatographic test (ict)
• elisa
• bovine rotavirus
• bovine coronavirus (bcv)