Synthetic Norovirus RNA Reference Materials Enable Quality Control Across Commercial Detection Kits
Synthetic norovirus GI and GII RNA reference materials were developed via in vitro transcription and characterised across five laboratories using four commercial real-time PCR assays. The materials were homogeneous, stable for one year at -20 degrees C or below, and compatible with all six commercial detection kits tested. These reference standards fill an important gap for quality assurance in norovirus molecular diagnostics given the virus's high sequence variability.
The original study
Development and Characterization of Synthetic Norovirus RNA for Use in Molecular Detection Methods.
- Authors
- Cho EJ, Cha Y, Lee SK, Kim HS, Kim JS, Lee EJ, et al.
- Journal
- Annals of laboratory medicine
- Type
- Journal Article, Multicenter Study
- PMID
- 36045055
Original abstract
BACKGROUND: Reference materials are essential for the quality assurance of molecular detection methods. We developed and characterized synthetic norovirus GI and GII RNA reference materials. METHODS: Norovirus GI and GII RNA sequences including the ORF1-ORF2 junction region were designed based on 1,495 reported norovirus sequences and synthesized via plasmid preparation and in vitro transcription. The synthetic norovirus GI and GII RNAs were evaluated using six commercial norovirus detection kits used in Korea and subjected to homogeneity and stability analyses. A multicenter study involving five laboratories and using four commercial real-time PCR norovirus detection assays was conducted for synthetic norovirus RNA characterization and uncertainty measurements. RESULTS: The synthetic norovirus GI and GII RNAs were positively detected using the six commercial norovirus detection kits and were homogeneous and stable for one year when stored at -20°C or -70°C. All data from the five laboratories were within a range of 1.0 log copies/μL difference for each RNA, and the overall mean concentrations for norovirus GI and GII RNAs were 7.90 log copies/μL and 6.96 log copies/μL, respectively. CONCLUSIONS: The synthetic norovirus GI and GII RNAs are adequate for quality control based on commercial molecular detection reagents for noroviruses with high sequence variability. The synthetic RNAs can be used as reference materials in norovirus molecular detection methods.