期刊
INTERVIROLOGY
卷 62, 期 3-4, 页码 145-155出版社
KARGER
DOI: 10.1159/000502823
关键词
Chikungunya virus; Envelope 1 antigen; Monoclonal antibody; Coumarin-derived dendrimer
类别
资金
- Priority Research Centers Program through the National Research Foundation of Korea (NRF) - Ministry of Education [NRF2015R1A6A1A03032236]
Background: When infected with the chikungunya virus (CHIKV), 3% to 28% of CHIKV-infected individuals remain asymptomatic, necessitating the development of improved high-throughput screening methods to overcome the limitations of molecular diagnostics or enzyme-linked immunosorbent assays (ELISAs). Objective: In this study, two novel monoclonal antibodies (mAbs) targeting envelope 1 (E1) of CHIKV were developed and applied in a fluorescence-linked immunosorbent assay (FLISA) using coumarin-derived dendrimer as the fluorophore. Methods: The performance of the FLISA was compared with that of ELISA. Results: Using the two novel mAbs (2B5 and 2C8), FLISA could detect 1 x 10(5) PFU/mL of CHIKV, exhibiting a 2-fold lower limit of detection (LOD) compared to ELISA. The LOD of FICT corresponded to a comparative threshold value of 23.95 and 4 x 10(6) of RNA copy number/mu L. In the presence of human sera and blood, virus detection by FLISA was 3-fold better than ELISA, with an LOD of 2 x 10(5) PFU/mL. Sera and blood interfered with the ELISA, resulting in 6 x 10(5) PFU/mL as the LOD. Conclusions: FLISA using two novel mAbs and coumarin-derived dendrimer is a superior diagnostic assay for detecting CHIKV in human sera and blood, compared to conventional ELISA.
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