Real-time PCR assay as a simple and efficient tool for viral stability study

Area of Science:

• Virology
• Molecular Biology
• Pharmaceutical Science

Background:

• Regulatory agencies require risk assessment for viruses released from patients in oncolytic virus trials.
• Evaluating viral shedding and stability in biological matrices like urine is critical for clinical sample management.

Purpose of the Study:

• To establish a real-time PCR method for assessing viral shedding and stability in human urine.
• To provide a simple, efficient, and high-throughput assay for viral stability testing.

Main Methods:

• Incubation of viral drug product in human urine.
• Application of real-time PCR to quantify viral levels and assess stability.
• Testing viral stability under various temperatures and detergent conditions.

Main Results:

• A real-time PCR method was successfully established for assessing viral shedding and stability in human urine.
• The method demonstrated efficiency and high throughput for analyzing a nonenveloped, single-stranded RNA virus.
• Viral stability information was obtained at different temperatures and in the presence of detergents.

Conclusions:

• The developed real-time PCR method offers a valuable tool for assessing viral stability in human urine.
• This approach is critical for the safe collection, transport, storage, and testing of clinical samples.
• The methodology may be applicable to other RNA viruses, including SARS-CoV-2.