Peptide aptamer-based time-resolved fluoroimmunoassay for CHIKV diagnosis

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Viral Immunology

Background:

• Chikungunya virus (CHIKV) and Dengue virus (DENV) share similar symptoms, leading to frequent misdiagnoses.
• A diagnostic system capable of differentiating between CHIKV and DENV is crucial for accurate patient management.

Purpose of the Study:

• To develop a novel peptide aptamer with high affinity and specificity for Chikungunya virus (CHIKV).
• To construct a peptide aptamer-based Time-Resolved Fluoroimmunoassay (TRFIA) for efficient CHIKV detection.

Main Methods:

• Computer-aided design and molecular docking were used to identify CHIKV-specific peptide aptamers (B2 and B3).
• A sandwich-Time-Resolved Fluoroimmunoassay (TRFIA) was constructed using selected peptide aptamers.
• The assay's performance was evaluated using CHIKV E2 protein in PBS, serum, and urine samples.

Main Results:

• Peptide aptamer B2 demonstrated highly specific detection of CHIKV E2 with a detection limit of 8.5 ng/mL in PBS.
• The assay showed acceptable performance in clinical specimens (serum and urine), with detection limits of 57.8 ng/mL and 147.3 ng/mL, respectively.
• A good linear correlation between fluorescent signal intensity and E2 protein concentration (0-1000 ng/mL) indicated potential for quantitative detection.

Conclusions:

• High-affinity and specific peptide aptamers are effective for rapid diagnostic element screening.
• The developed peptide aptamer B2 offers superior detection of CHIKV viral particles compared to traditional antibodies.
• This peptide aptamer-based TRFIA assay provides a promising tool for accurate CHIKV diagnosis.