Integrated CRISPR-Cas12aand RAA one-pot visualstrategy for the rapid identification of Streptococcusequi subspecies equi

We specifically designed RAA primers and CRISPR RNA totarget the eqbE gene of S.equi, and we have carefully optimized the reaction systems for this purpose. The newly established visual diagnostic method hasshown to be highly effective, demonstrating 97.14% specificity and 100%sensitivity, with the capability to detect as few as 5.6×100 copies of the target.This is the first study to propose the combined application of RAA and CRISPR/Cas12a for the on-site rapid detection of S.equi. This is the first study to propose the combined application of RAA and CRISPR/Cas12a for the on-site rapid detection of S.equi, which enables visual point-of-care diagnosis of Strangles.