Development of Standard and Rapid Antigen Detection Assays for Human Metapneumovirus

ABSTRACT Human metapneumovirus (hMPV), identified as a novel respiratory pathogen in 2001, is responsible for causing acute respiratory illness across various patient demographics. Early detection of hMPV is crucial for administering timely treatment, thereby controlling the virus's propagation. There is a pressing need for the development of a more convenient and expeditious detection strategy for hMPV. The present study focused on the expression and purification of the highly conserved nucleoprotein (N) of hMPV, which served as an antigen in the generation of specific nanobodies and mouse monoclonal antibodies. Subsequently, we evaluated the efficacy of these immunological reagents in detecting the hMPV antigen through the application of double‐antibody sandwich enzyme‐linked immunosorbent assay and colloidal gold lateral flow chromatography test strips. These detection methods were successfully utilized on the recombinant antigen, cell culture‐derived hMPV, and nasopharyngeal swab specimens. The findings offer promising avenues for the development of convenient and rapid detection techniques, which are particularly pertinent during the virus's epidemic seasons.