Transcriptomics as an Adventitious Virus Detection Strategy
As an analytical platform, the Sanofi Pasteur Analytical Sciences (Toronto) Molecular Biology Centre (MBC) is applying and optimizing the use of high-throughput sequencing (HTS). The detection of adventitious viruses in biological products relies on a set of methods defined by the regulatory agencies called the compendial methods. High-throughput sequencing has the potential to improve the current breadth of detection and to remove some poorly performing compendial in vivo animal tests.
Since 2016, the Molecular Biology Centre has used HTS for the detection of adventitious viruses. An unbiased nucleic acid extraction procedure was optimized and published in 2018 by Sanofi Pasteur that can capture viral nucleic acids from a wide variety of viruses with different physical and biochemical properties. In addition to the basic extraction, a 2017 Mitacs collaboration with York University evaluated and incorporated the use of ribosomal RNA depletion and an optimized cDNA synthesis step into the sample preparation protocol.
The developed sample extraction pipeline uses a total genomics approach where all nucleic acids within a sample are extracted and sequenced. One drawback of this approach is that it detects only viral nucleic acids and cannot distinguish between infectious viral particles and non-infectious viral nucleic acids (...)