Next Generation Sequencing (NGS) Internal Controls for Adventitious Agent Testing to Ensure Sensitivity for All Targets in Every Sample
Our proposal will develop a test kit—AccuKit— that streamline adventitious agent screening while directly ensuring every target is measured with the required sensitivity.
Categories
Drug substance
Drug product
Assays
Project status
100% Completed
Industry Need
Adventitious agent testing of Master and Working cell banks is one critical aspect of purity testing as it minimizes risk of upstream contamination.
Adventitious agent testing is moving away from the use of animal models to more in vitro tests such as cell culture and PCR-based methods.
Solution
AccuGenomics and NC State will develop a test kit (“AccuKit”) that will streamline adventitious agent screening while directly ensuring every target is measured with the required sensitivity.
AccuKits will incorporate PCR primers specific for 22 known adventitious viruses and bacteria for high throughput NGS screening assays.
The kit will contain a mixture of competitive templates for every adventitious agent spiked into every sample as internal sensitivity controls; the expected test result for contamination should be positive for every spike-in target and negative for the native targets.
The platform allows for additional primer sets to be added to the AccuKit to meet future adventitious agent detection needs. The team intends to advance its MRL-4 level technology to MRL-6.
Outputs/Deliverables
Demonstrated new utility of AccuGenomics SNAQ technology as a QC tool to ensure every NGS panel target for every sample met the required test sensitivity requirement.
Created a fast (1 day) and sensitive (10 copy LOD) targeted NGS panel as a test bed for SNAQ QC technology.
Additional Project Information (Members Only)
Login to the NIIMBL member portal to access more, including: