Novaplex sars cov 2 variants 7 assay

A retrospective cohort of SARS-CoV-2 reinfection cases was carried out at the University Hospital of Verona from 24 February to 10 August 2020, 2022, among 7638 HW, within a dynamic cohort [6 (link)] included in the ORCHESTRA project, a 15 countries multi-centre study.
Reinfections were identified by screening and contact tracing, including those that occurred >90 days after the prior infection and if new COVID-19 symptoms began after the resolution of prior symptoms [7 (link)]. Only HW having multiple SARS-CoV-2 infections with thrice-positive swabs were included. Patient, infection, and virus characteristics were collected.
SARS-CoV-2 genome detection was performed on nasopharyngeal swabs via RT-PCRs. Samples previously tested positive were analysed to assess the SARS-CoV-2 variant by Novaplex™ SARS-CoV-2 Variants VII Assay (Seegene, Seoul, South Korea).

The Novaplex SARS-CoV-2 Variants VII Assay (Novaplex) (Seegene, Seoul, Republic of Korea) was carried out according to the manufacturer’s instructions using the CFX96 Real Time PCR Detection System and the BioRad CFX96 Manager software (Bio-Rad, Hercules, CA, USA). The results were analyzed via automated data interpretation using Seegene Viewer software. The Novaplex assay is based on MuDT™ technology, which detects multiple targets with individual Ct values in a single channel without melting curve analysis. By utilizing the change in fluorescence signals between two different temperatures of detection, MuDT™ provides the “real” Ct value of each pathogen even in co-infected cases. MuDT™ allows for the genotyping of four SNP targets in a single tube. The set of primers can detect three SARS-CoV-2 variants of the S gene showing ∆69/70, E484A, and N501Y (Figure 1). The RdRP gene was considered as an internal control. All the NPSs that were only positive for the RdRP gene were classified as positive, but undetermined. Samples showing ∆69/70 deletion (Δ69/70) and N501Y were considered positive for the Alpha variant. Samples with ∆69/70, E484A, and N501Y SNPs were assigned to the Omicron BA.1 variant, while E484A and N501Y SNPs were attributed to the Omicron BA.2 variant.