Microchemical Journal.
2025 Jun
DOI:
10.1016/j.microc.2025.113779
Jiayi Yang, Mingwei Liu, Yunhua Gao, Huijie Li, Lianhua Dong
Highlights
• The RT-dPCR method was established to accurately quantify the norovirus GⅡ.4 pseudovirus RM.
• The norovirus GⅡ.4 pseudovirus RM containing ORF2 RNA exhibits good homogeneity and stability.
• The value-assigned pseudovirus RM can serve as a reliable quality control tool for norovirus GⅡ.4 detection assays.
Abstract
The human norovirus (NV) is widely recognized as a pathogen causing acute gastroenteritis, particularly associated with elevated morbidity and mortality rates in pediatric and geriatric populations. While current detection methods, such as those based on immune reactions and quantitative polymerase chain reaction (qPCR), have been developed by researchers, these techniques do not provide absolute quantification. In this study, we established a reference material (RM) comprising pseudovirus particles containing NV ORF2 RNA. The RM was quantified using reverse transcription digital PCR (RT-dPCR), demonstrating ideal homogeneity and stability. The assigned value of the RM, including expanded uncertainty, was determined to be (3.42 ± 0.69) × 10³ copies/μL. This RM can be effectively utilized to validate commercial diagnostic kits and platforms, thereby enhancing the accuracy and reliability of acute gastroenteritis diagnosis.
Tags:
Human Norovirus GⅡ.4; Acute Gastroenteritis; RT-dPCR; Reference Material