Calibration of a suitable enzyme-linked immunosorbent assay (ELISA) for Nilaparvata lugens (stal) antigen detection

Abstract

Direct, indirect and double antibody sandwich methods of enzyme-linked immunosorbent assay (ELISA) were examined with the monoclonal antibodies against Nilaparvata lugens (Stal), 3B2 and 4B8, for their ability to detect the antigen of N. lugens in the gut of predator. The results showed the non-target antigen (predator) in a sample had a great effect on ELISA tests. The direct ELISA and indirect ELISA were affected more than the double antibody sandwich ELISA. The sandwich ELISA was the most sensitive, capable of detecting down to about 16.90 ng/mL (1/17509 female) compared with 67.59 ng/mL (1/4378 individual) for the indirect ELISA and 135.18 ng/mL (1/2189 individual) for the direct ELISA. We established the double antibody sandwich methods of ELISA with the 4B8 diluted 1000 times (28.13 μg/mL）, enzyme-linked antibody, HRP-4B8, diluted 4000 times (1.04 μg/mL）and sample diluted 100 times for N. lugens antigen detection. The negative effects of the non-target antigens were lower than 5% and 1.69 μg/mL prey antigens (1/175 individual of the female of N. lugens） remained in a predator gut could be detected under this system.

Keywords

• <italic>Nilaparvata lugens</italic>
• monoclonal antibody
• ELISA
• sensitivity
• non-target antigen