Preparation of Fluorescent Antibody Conjugate in Goats

Purification of ribonucleoprotein (RNP) from neuroblastoma-infected cells with a street rabies virus (SRV), in this case canid RABV, or a fixed strain (such as ERA) and a member of phylogroup II and lyssaviruses found exclusively on the African continent (e.g. Mokola virus, MOKV), is described. The RNP is purified by employing ultracentrifugation in a cesium chloride (CsCl) gradient, and the resulting product checked for intactness and authenticity by Western blotting and immunoblotting with a specific lyssavirus antibody. The protein product can then be used for immunizing goats to generate polyclonal antibodies (PAbs) for labeling with either biotin for use in the direct rapid immunohistochemical test (dRIT) or fluorescein isothiocyanate (FITC) for lyssavirus antigen detection in the direct fluorescent antibody test (dFAT).