Visual detection and identification of influenza A viruses by nucleic acid probe‐enabled lateral flow assay

Diagnosis of influenza A viral infection is crucial for preventing disease transmission and providing effective clinical treatments. There is an increasing need for convenient detection methods to enable simple yet precise identification of viral infections. Here, we developed a nucleic acid probe‐enabled lateral flow assay (NALFA) to realize visual detection and identification of influenza A viral infections (H1N1 and H3N2) of high sensitivity and specificity. Viral RNA targets were recognized by a padlock probe, which was circularized to induce rolling circle amplification (RCA). RCA products were enzymatically cleaved into short amplicons to complex with capture DNA probes for gold colloidal‐induced visual lateral flow assay. NALFA achieved attomolar (aM) sensitivity for both standard viral RNAs, along with high specificity. While applying clinical samples (16 H1N1 patients, 12 healthy controls), NALFA exhibited high detection accuracy to successfully discriminate infected samples from noninfected samples. NALFA represents a potent and convenient nucleic acid detection assay that shall find its applications in fields of viral detection and beyond.