The invention discloses a
CRISPR-Cas12a (clustered regularly interspaced short palindromic repeats-associated 12a)-based
rapid detection method for an OsHV-1 (OsHV-1)
virus The method comprises the following steps: (1) RPA amplification: amplifying a
target gene segment by using the primer in claim 1 and taking
DNA of
oyster tissues as a template; (2) obtaining ORF38-crRNA (
Ribose Nucleic Acid), wherein the ORF38-crRNA is UCCUUGCCUCCAAGGUUCUUAUUACUUAUUAGUGAGUCGUUUUA, and the ORF38-crRNA is obtained by the following steps of: (1) obtaining the ORF38-crRNA; (2) obtaining the ORF38-crRNA, and (3) detecting whether a
specific fluorescence signal is generated on the
target gene segment by using a
CRISPR-Cas12a detection
system, if the
specific fluorescence signal is generated on the
target gene segment, indicating that the
oyster tissue contains the OsHV-1
virus, and if the
specific fluorescence signal is not generated on the target
gene segment, indicating that the
oyster tissue does not contain the OsHV-1
virus. According to the method, the trans-cleavage activity of RPA and
CRISPR-Cas12a is combined, an FAM labeled ssDNA
probe signal is adopted for detection, the sensitivity is high, the specificity is high, the
detection limit can reach 1 copy / mu L, and the detection time does not exceed 40 minutes. The method can be widely applied to the fields of
aquaculture disease monitoring, aquatic
food safety detection, environment monitoring and the like.