Evaluation of the RHAM-based Pluslife Rapid SARS-CoV-2 assay and comparison with EasyNAT Rapid SARS-CoV-2 assay and Wondfo 2019-nCoV antigen assay

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains a significant public health challenge. This study evaluated the Pluslife SARS-CoV-2 assay employing proprietary RNase Hybridization-Assisted amplification (RHAM) isothermal amplification technology, with comparative performance assessment against the EasyNAT Rapid SARS-CoV-2 assay and the Wondfo 2019-nCoV antigen assay. Analytical testing determined the Pluslife assay's limit of detection to be 400 copies/mL, with no cross-reactivity against common respiratory viruses and no interference from tested substances. Using RT-qPCR as the reference standard on 197 nasopharyngeal swabs, the Pluslife assay showed a positive percentage agreement (PPA) of 96.30% and a negative percentage agreement (NPA) of 99.30% (<i>κ</i> = 0.962, <i>P</i> < 0.001). The EasyNAT assay demonstrated a PPA of 96.30% and NPA of 98.60% (<i>κ</i> = 0.949, <i>P</i> < 0.001). The Wondfo antigen assay showed a PPA of 81.48% and NPA of 100% (<i>κ</i> = 0.865, <i>P</i> < 0.001). The Pluslife assay demonstrates comparable diagnostic accuracy to EasyNAT for SARS-CoV-2 detection, while offering advantages in portability, speed, and cost. Its performance significantly exceeded that of the Wondfo antigen assay, particularly in reducing false negatives. The Pluslife Mini Dock platform shows potential for rapid SARS-CoV-2 screening in diverse settings.IMPORTANCEThe newly developed Pluslife Rapid severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) assay is a next-generation molecular point-of-care testing (POCT) method, based on RNase Hybridization-Assisted amplification (RHAM), a self-developed isothermal amplification molecular detection technology of Pluslife Biotech. The purpose of this study is to evaluate the analytical and clinical performance of this new SARS-CoV-2 POCT method and compare the clinical performance with the EasyNAT Rapid SARS-CoV-2 assay and the Wondfo 2019-nCoV antigen assay. The results of this study are of great significance to clinical practice.