Study design: Assay optimisation. Methods: Four sgRNA between 20 and 24 nucleotides in length complementary to different regions of the segment 7 (M gene) of H3N8 EIV, were designed using the CRISPRscan software, based in CAS12a predicted guides. Complete M gene of EIV H3N8 Florida clade 1 (FC1) and Florida clade 2 (FC2) representative strains were amplified by one step RT-PCR. The CRISPR-Cas12 detection step was carried out by incubating the PCR products with the CRISPR complex at 37ºC, and fluorescence was measured for up to 60 minutes every 5 minutes (ssDNA FQ substrate λex = 485 nm; λem = 535 nm). Results: All the designed sgRNA were able to detect the EIV H3N8 of FC1 while 2 of them detected FC2 representative strains, between 5 and 20 minutes. Main limitations: Limit of detection was not established and clinical samples were not assayed. Conclusions: CRISPR-Cas12a technology seems to be a promising system for detection of equine influenza virus, offering a convenient tool for low-resource laboratories. Ethical animal research: Not applicable.