Biosafety Open Forum

Siemon described how Sanofi Pasteur’s Analytical Sciences department has been applying High-throughput sequencing technology for the detection of adventitious viruses since 2011. High-throughput sequencing (HTS) is capable of detecting both known and unknown adventitious viruses that may be present in a sample. To demonstrate equivalency with existing tests, Sanofi Pasteur evaluated the detection sensitivity of HTS using a well-characterized panel of model viruses and determined the limit of detection within a viral seed matrix, and validated the assay within a GMP environment.

Thanks to Blandine de Saint Vis, we gained insight into the viral purity testing approaches undertaken at Boehringer Ingelheim Animal Health (BIAH), striving to produce vaccines for animals including livestock and pets. A thorough analysis for viral contaminations in seed and starting material is crucial. Blandine shared an overview of the adventitious agent detection process and the objectives behind the BIAH’s motivation to embrace new approaches such as NGS. While sharing the benefits of NGS such as its speed, cost-effectiveness, and ability to detect all potential contaminants, Blandine also explained the complexities associated with the data analysis, validation, and determining criteria to define a true contaminant. To conclude, Blandine explained how BIAH validates NGS-based assays using a spike-in study and a panel of model viruses. She described the need to assess the reference database used and optimize the bioinformatics pipeline. Her team is currently using Genedata Selector’s bioinformatics pipeline for NGS-based data analysis. 

This presentation expanded the understanding of the analysis requirements of successful adventitious agent detection. Starting with the structure of a basic pipeline, Devon shared other approaches to improve the sensitivity and specificity of an NGS-based biosafety assay. He also discussed the impact of the selected reference viral database (RefSeq or C-RVDB) on the results obtained. Further studies performed by the scientific consultant team of Genedata identified the likely sources of false positives demonstrating the need to sequence the host cell line genome. Devon concluded by explaining how Genedata Selector simplifies the data processing and analysis of NGS-based biosafety assay data while allowing rapid data query for efficient decision-making. The automation and easy reporting capabilities of the software also enable companies to obtain the documentation they need for regulatory compliance.