Abstract: : Objective To establish a luciferase immunosorbent assay (DENV-LISA) based on dengue virus (DENV) E protein, a specific antigen of DENV, for detection of DENV IgG antibody. Methods The fused expression plasmids of DENV1-E1 and DENV2-E2 with luciferase were constructed. The plasmids were transfected into 293T cells, and the fusion protein containing the specific antigen and luciferase was obtained for establishing DENV-LISA. The specificity and sensitivity of DENV-LISA were assessed and compared with those of commercial DENV IgG antibody detection kit (ELISA). Results The established DENV-LISA had a positive detection rate of 32.4% and a specificity of 96.6%, showing a similar positive detection rate with the commercial ELISA kit (35.3%; P>0.05). DENV-LISA was capable of detecting positive samples with a 1: 6400 dilution with a high sensitivity. The test values of DENV-LISA did not differ significantly between plates or within plates in the same batch (P> 0.05), suggesting a good reproducibility of the test. Conclusion The luciferase immunosorbent assay based on DENV E protein has high specificity and sensitivity for detecting DENV IgG antibody, and can be used for early screening, surveillance and epidemiological investigation of DENV infection.

Key words: dengue virus; luciferase; E protein; IgG antibody; detection