Tripropylamine facilitates efficient preparation of flower-like nanozyme for rapid and sensitive Mpox point-of-care testing

Recurrent outbreaks of Mpox have highlighted the importance of rapid diagnosis to effectively control virus transmission from infected individuals. Traditional laboratory-based detection methods are time-consuming, complex, and unsuitable for point-of-care testing. Therefore, we developed a lateral flow immunoassay (LFIA) using platinum–gold nanoflowers (PtAu NFs) as a labeling material, and this technique was integrated with recombinase polymerase amplification (RPA) to detect Mpox within 25 min. From material preparation to detection, the speed and simplicity of detection were comprehensively optimized in this study. Moreover, a novel material preparation method based on tripropylamine (TPrA) as a reducing agent for the rapid preparation of PtAu NFs has been demonstrated. Unlike traditional reducing agents, this method does not require high temperatures and extended incubation periods, thereby providing a novel strategy for preparing nanomaterials. Furthermore, the traditional RPA–LFIA strategy was improved by directly modifying the primer label, avoiding the use of probes and exonucleases, which saves reagent costs. Under optimized conditions, the RPA–PtAu–LFIA platform had a visual limit of detection of 1 copy/μL, with high specificity for Mpox and other poxviruses. Furthermore, the sensitivity of this platform was validated using 29 different clinical samples obtained from the pustules, saliva, anus, and throat of infected individuals; the results obtained using this platform were comparable to those obtained using quantitative polymerase chain reaction. This study provides an excellent platform for preparing nanomaterials and detecting Mpox.