M13mp18 nanoscaffold-based strip sensor for detecting influenza A virus (H1N1)

We report the development of an amplification-free nucleic acid assay using lateral flow test strips. M13mp18 nanoscaffold signal amplification permitted the visual detection of H1N1 RNA in 20 min. M13mp18 nanoscaffold supported a set of backbone, variable, detection, and modified oligonucleotides. A bridging-type FAM probe/H1N1 RNA/M13mp18 was used in the lateral flow test strip format. The bridging-type complexes were captured on the test zone. The lower limit of detection of specific H1N1 RNA was 1 pM. The method directly detected H1N1 RNA without amplification, eliminating the risk of amplicon contamination and lessening the chance of false positives. This biosensor has potential value for field testing in resource-limited settings and monitoring viral loads in recovering patients. The amplification-free lateral flow detection of RNA provides an alternative between amplification-based RNA assays and protein antigen assays for facilitating the rapid identification of H1N1 virus during point-of-care testing.