Journal of Biomedical Nanotechnology · 2019 · 109 citations · 0 references
Life SciencesBiotechnologyMolecular BiologyMolecular BiotechnologyNanobiotechnologyVirologyCrispr-cas SystemsDna NanotechnologyInfectious Disease Molecular TestingRna EditingRna StructureRna-based DiagnosticsAvian Influenza DiagnosisAvian Influenza VirusCas Nuclease SpecificityCrispr-based Molecular DiagnosticsCrispr-cas13 SystemsSingle-guide Rna Design
transcribed RNA of H7N9 HA gene has been used as a target, to design a specific crRNA. The activity of Cas13a was verified with a single-stranded RNA-bound fluorescent group and a quenching fluorophore as signals. Using Cas13a, a room temperature H7N9 detection system was established. Detection of 1 nm of single-stranded RNA can be done within 5 min. When combined with the RT-RPA and T7 transcription system at room temperature, the detection limits of HA and NA are 1 fM and the reaction time is 50 min. Excellent specificity was achieved by comparison with subtype viruses such as H1N1 and H5N1. The rapid detection method based on CRISPR-Cas13a nanomachine H7N9 has been successfully established, which can detect H7N9 quickly and specifically. In the future, it can be quickly detected in the field with portable fluorescence detector.