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Updated: May 11, 2026
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Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
Published on: May 26, 2013
Virus detection by short read high throughput sequencing in a high virus low cellular background
Pei-Ju Chin1, Christophe Lambert2, Pascale Beurdelay3,4
1Office of Vaccines Research and Review, Center for Biologics Evaluation and Review, U.S. Food and Drug Administration, Silver Spring, MD, 20993, USA.
NPJ Vaccines
|March 29, 2025
Summary
High-throughput sequencing (HTS) effectively detects adventitious viruses in biological products. Optimizing HTS workflows enhances sensitivity for viral safety testing, ensuring product safety.
Area of Science:
- Virology
- Genomics
- Biotechnology
Background:
- Biological product safety relies on detecting adventitious viruses.
- High-throughput sequencing (HTS) offers a sensitive method for viral detection.
Purpose of the Study:
- To evaluate the sensitivity and breadth of HTS for adventitious virus detection.
- To validate HTS protocols for viral safety testing of biologicals.
Main Methods:
- Seven laboratories assessed HTS using five reference viruses (EBV, FeLV, RSV, Reo1, PCV1).
- Viruses were spiked into adenovirus at concentrations from 10^2 to 10^4 GC/mL.
- Targeted and non-targeted bioinformatic analyses were employed.
Main Results:
- All laboratories detected viruses at 10^4 GC/mL using both analysis types.
- At 10^3 GC/mL, targeted analysis detected all five viruses; non-targeted analysis detected Reo1 and EBV.
- Some labs achieved detection limits of ≤10^2 GC/mL.
Conclusions:
- HTS is a viable method for adventitious virus detection in biological manufacturing.
- Optimization of HTS workflows is crucial for improving detection limits.
- This study provides a framework for HTS validation in viral safety testing.
