Rapid and visual detection of enterovirus using recombinase polymerase amplification combined with lateral flow strips

Enteroviruses (EVs) are the most common causative pathogen of infection in children aged under 5 years. Routine laboratory methods for detecting EV are time consuming, labor intensive, and require sophisticated thermal cycling instruments and skilled operators, which are not available in limited-resource settings. In this study, a novel isothermal amplification, recombinase polymerase amplification (RPA) combined with lateral flow strips (LFS), was established to detect EVs. Specific RPA-LFS primers and probe were designed to target the highly conserved regions of 5'-UTR. The analytical sensitivity for detection of EV was 5 copies per reaction, with 100 % specificity. The clinical performance was evaluated using 177 clinical samples, and the coincidence rates between RPA and commercial quantitative real-time PCR was 100 %. In conclusion, the RPA-LFS developed in this study is a rapid, specific, sensitive, and accurate assay for detecting EV and could thus be an ideal diagnostic tool for EV infections in limited-resource settings. It is the first time that RPA-LFS assay has been applied to the detection of EV infection.