Fluorescent Immunosorbent Assay for Chikungunya Virus Detection

Area of Science:

• Virology
• Immunology
• Biotechnology

Background:

• Chikungunya virus (CHIKV) infection presents diagnostic challenges due to asymptomatic cases (3-28%).
• Current molecular diagnostics and enzyme-linked immunosorbent assays (ELISAs) have limitations for high-throughput screening.

Purpose of the Study:

• To develop and evaluate novel monoclonal antibodies (mAbs) against CHIKV envelope 1 (E1).
• To establish a fluorescence-linked immunosorbent assay (FLISA) utilizing these mAbs and a coumarin-derived dendrimer fluorophore for CHIKV detection.

Main Methods:

• Development of two novel mAbs (2B5 and 2C8) targeting CHIKV E1.
• Application of these mAbs in a FLISA format.
• Comparative performance analysis of FLISA against conventional ELISA using human sera and blood samples.

Main Results:

• FLISA achieved a 2-fold lower limit of detection (LOD) for CHIKV compared to ELISA (1 × 10^5 PFU/mL).
• FLISA demonstrated 3-fold better virus detection in human sera and blood than ELISA.
• FLISA exhibited an LOD of 2 × 10^5 PFU/mL in complex biological matrices, whereas ELISA's LOD was 6 × 10^5 PFU/mL due to interference.

Conclusions:

• FLISA employing novel mAbs and a coumarin-derived dendrimer is a highly effective diagnostic tool for CHIKV.
• This FLISA method surpasses conventional ELISA in sensitivity and performance for detecting CHIKV in human sera and blood.