Peptide aptamer-based time-resolved fluoroimmunoassay for CHIKV diagnosis

• Published in: Virology journal Vol. 20; no. 1; p. 166
• Main Authors: Liu, Tonggong, Gao, Cheng, Wang, Jingzhe, Song, Jianning, Chen, Xi, Chen, Hongfang, Zhao, Xiaona, Tang, Huanwen, Gu, Dayong
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 27.07.2023; BioMed Central; BMC
• Subjects: Affinity; Antigens; Aptamers; Chikungunya fever; Chikungunya Virus; Computer-aided design; Computer-aided medical diagnosis; Dengue fever; Diagnosis; E2 protein; Epidemics; Genomes; Health aspects; Hydrogen bonding; Immunoassay; Immunology; Infections; Methods; Molecular docking; Monoclonal antibodies; Mosquitoes; Nanoparticles; Peptide aptamer; Peptides; Polyclonal antibodies; Proteins; Reagents; Serology; TRFIA; Viruses; Zika virus; China; Asia; United States > US; Chikungunya Virus; Computer-aided design; Peptide aptamer; TRFIA; Molecular docking
• ISSN: 1743-422X; 1743-422X
• DOI: 10.1186/s12985-023-02132-w

Chikungunya virus (CHIKV) and Dengue virus (DENV) have similar clinical symptoms, which often induce misdiagnoses. Therefore, an antigen detection diagnostic system that can clearly identify these two viruses is desirable. In this study, we developed a novel peptide with high affinity and specificity to CHIKV, and further constructed peptide aptamer-based TRFIA assay to efficiently detect CHIKV. Peptide aptamer B2 (ITPQSSTTEAEL) and B3 (DTQGSNWI) were obtained through computer-aided design and selected as CHIKV-specific peptide aptamers based on their high binding affinity, strong hydrogen bonding, and RMSD of molecular docking. Then, a sandwich-Time-Resolved Fluoroimmunoassay (TRFIA) was successfully constructed for the detection of the interaction between peptide aptamers and viruses. When using B2 as the detection element, highly specific detection of CHIKV E2 was achieved with detection limits of 8.5 ng/ml in PBS solution. Variation coefficient between inter-assay showed the disturbances received from the detection of clinical fluid specimens (including serum and urine), were also within acceptable limits. The detection limits for 10-fold dilution serum and urine were 57.8 ng/mL and 147.3 ng/mL, respectively. The fluorescent signal intensity exhibited a good linear correlation with E2 protein concentration in the range of 0-1000 ng/mL, indicating the potential for quantitative detection of E2 protein. These results demonstrate that the construction of peptide aptamers with high affinity and specificity provides an excellent method for rapid diagnostic element screening, and the developed peptide aptamer B2 contributed to better detection of CHIKV viral particles compared to traditional antibodies.