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

ABSTRACT:

Background/objectives

Novel influenza A viruses (NIAV) infection is a zoonotic acute respiratory disease. 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.

Methods

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 verifie

SUBMITTER: Luo Z 

PROVIDER: S-EPMC13623710 | biostudies-literature | 2026

REPOSITORIES: biostudies-literature