Development of an ELISA to quantitatively determine the nucleoprotein of rabies virus / 临床检验杂志

ABSTRACT

Objective To develop a double-antibody sandwich ELISA for determining the concentration of nucleoprotein (NP) of rabies virus in various products of rabies vaccine.Methods The purified rabies antibodies from a rabbit were used to coat microwell plates. Horseradish peroxidase-conjugated anti-NP monoclonal antibody was used to probe the NP bound to coated antibodies.The assay was used to quantitatively determine the concentration of NP of rabies virus.Results The results showed that the coefficient of linear correlation was higher than 0.97.The optimal linear range was 0.000625~0.01 IU/ml and the detection limit was 0.000625 IU/ml. The recovery rate was 102~109% and the coefficient of variation was only 7.2%~9.4%.No cross reactions were observed with bovine serum,bovine serum albumin,ovalbumin,refined solution of influenza vaccine,encephalitis B vaccine,and hepatitis A vaccine.Conclusions The results indicated that the assay is specific,sensitive,accurate,reproducible,and stable,and could be suitable for quantitative determination of different rabies vaccine's processes products.