Application of monoclonal antibody DSSC7 for detecting dengue infection in Aedes aegypti based on immunocytochemical streptavidin biotin peroxidase complex assay (ISBPC).
AbstractAedes aegypti mosquito is the important vector of dengue fever and chikungunya fever. Therefore, forvirus detection in the mosquito, the possibility of cross-reactivity with chikungunya virus must beconsidered. The laboratory studies were aimed at characterizing the monoclonal antibody DSSC7,and its application for detecting dengue (DENV) antigen on the various organs of orally-infected Ae.aegypti in the paraffin-embedded tissue sections, viz. head squash, abdomen squash, based on immunocytochemical streptavidin-biotin-peroxidase complex (ISBPC) assay. Determination of the antibody class and subclass was based on antigen-mediated ELISA (enzyme-linked immunosorbentassay). The specificity of monoclonal antibody DSSC7 was determined by Western blotting method,using DENV-1, DENV-2, DENV-3, DENV-4, and chikungunya antigen. The presence of DENV antigen inthe various organs of the orally-infected Ae. aegypti were microscopically optimized in the paraffin-embedded tissue section using ISBPC assay and monoclonal antibody DSSC7 (diluted 1:10, 1:50,1:100 in phosphate buffer saline) as a primary antibody. The specificity of the immunocytochemical procedure is validated by negative controls and by positive controls that show that the antibody isbinding to an appropriate structure. The monoclonal antibody DSSC7 recognize DENV complexspecific and does not recognize chikungunya antigen. The monoclonal antibody belongs to IgG class,IgG1 subclass, and it is used as primary antibody to detect DENV infection in Ae. aegypti on tissue section in experimental infection based on ISBPC method. The infection rates of abdomen squash and head squash after incubation period of five days were 75% and 33.33% respectively.
Sutaryo, Umniyati SR, Artama WT, Wahyono D, Soeyoko, Mardihusodo SJ & Utoro T, Mulyaningsih B. (2008). Application of monoclonal antibody DSSC7 for detecting dengue infection in Aedes aegypti based on immunocytochemical streptavidin biotin peroxidase complex assay (ISBPC).. WHO Regional Office for South-East Asia.. http://www.who.int/iris/handle/10665/170473