Improved RSV Neutralization Assay Using Recombinant RSV Expressing Reporter Fluorescent Protein

Area of Science:

• Virology
• Immunology
• Assay Development

Background:

• Human respiratory syncytial virus (RSV) causes significant global mortality, particularly in young children and older adults.
• RSV neutralization assays are crucial for quantifying antibodies that confer protection against infection and severe disease.
• Existing assays often face limitations in speed, cost, and resource requirements.

Purpose of the Study:

• To develop an improved, simple, and economical RSV neutralization assay.
• To utilize a recombinant RSV (rRSV) expressing a reporter fluorescent protein for measuring viral growth.
• To enable efficient antibody titration for RSV surveillance and antiviral drug testing.

Main Methods:

• Developed a novel RSV neutralization assay using rRSV engineered to express a reporter fluorescent protein.
• Optimized cell culture conditions, determining 3 days of incubation to be sufficient for robust fluorescent signal detection.
• Utilized white 96-well plates for enhanced reporter activity stability compared to black plates.
• Validated the assay's performance against established methods like rRSV-Luciferase and ELISA.

Main Results:

• The assay successfully measured RSV growth via reporter activity using a standard plate reader.
• Sufficient fluorescent activity was achieved with low amounts of rRSV after 3 days of cell culture.
• White 96-well plates provided more stable reporter activity than black plates.
• The fluorescent reporter assay demonstrated comparable antibody titer detection capacity to existing methods.

Conclusions:

• The developed assay offers a simple, economical, and effective alternative for RSV neutralization testing.
• This method is suitable for high-throughput antibody titration, supporting RSV surveillance and antiviral research.
• The use of fluorescent reporter proteins provides a viable strategy for improving diagnostic assay efficiency.