The invention discloses a kit, application and method for rapidly detecting
mycoplasma synoviae based on an ERA-
CRISPR / Cas12a one-
step method, according to the
visual detection kit, on the basis of a new
nucleic acid detection target scpA of the
mycoplasma synoviae, a sample to be detected is split by a splitting buffer to release
genome DNA, then ERA-
CRISPR / Cas12a one-pot reaction is carried out in a constant-temperature water bath kettle or a
metal bath, and the sample to be detected is subjected to one-pot reaction, so that the ERA-
CRISPR / Cas12a one-pot reaction is carried out to rapidly detect the
mycoplasma synoviae through the ERA-CRISPR / Cas12a one-pot reaction. Under the mediation of a crRNA guide sequence, after Lb Cas12a
enzyme is combined with crRNA and target
DNA to form a
ternary complex, the trans-shear activity of cas12a is activated, an ssDNA fluorescent probe in a
reaction system can be arbitrarily
cut to release a fluorescent group on the probe, and an
ultraviolet lamp is used for detecting a fluorescent
signal or a
colloidal gold test strip is used for detecting whether FAM and
Biotin groups are
cut off for result judgment. According to the ERA-CRISPR / cas12a one-pot method technology for rapidly detecting the
mycoplasma synoviae, the
DNA target of the
mycoplasma synoviae is recognized and
cut through ERA amplification and CRISPR / cas12a specificity, the specificity is high, the detection sensitivity is as low as 3.81 pg / mu L, and the ERA-CRISPR / cas12a one-pot method technology can be used for rapidly detecting the
mycoplasma synoviae in clinical samples on site.