Table 1_Establishment and preliminary evaluation of an RT-ERA combined with CRISPR/Cas12a assay for detection of H5 and H7 influenza viruses.docx

The H5N1 and H7N9 subtypes cause high mortality, posing major public health threats. This study aimed to establish rapid, field-deployable subtype-specific nucleic acid detection assays for H5 and H7 by integrating reverse-transcription enzymatic recombinase amplification (RT-ERA) with CRISPR/Cas12a.

Two separate reactions are required per sample.</p>Methods<p>Conserved hemagglutinin (H) sequences were retrieved from the NCBI database, and primers, probes, and crRNAs were designed using Primer Premier 5 and Primer-BLAST. The sensitivity of RT-ERA alone was compared with that of the ERA-CRISPR/Cas12a using serially diluted virus strain nucleic acid. The specificity was verified against other respiratory pathogens with similar clinical manifestations.</p>Results<p>The optimal primers were F1R2 for H5 and F2R3 for H7.