The invention relates to the technical field of
plant virus detection, in particular to a pepper
vein flavivirus detection method and
system based on RT-RAA-
CRISPR / Cas12a and application of the pepper
vein flavivirus detection method and
system. The invention discloses a pepper
vein flavivirus detection method based on RT-RAA-
CRISPR / Cas12a. The pepper vein flavivirus detection method comprises the following steps: S1, extracting
virus RNA (Ribonucleic Acid) from a
plant leaf to be detected; s2, adding
RNA (Ribonucleic Acid) of a sample to be detected into the RT-RAA amplification
system, and carrying out isothermal amplification at 37-42 DEG C for 10-30 minutes to obtain an RT-RAA amplification product; s3, adding the RT-RAA amplification product into a
CRISPR / Cas12a (clustered regularly interspaced short palindromic repeats / CRISPR associated 12a) detection system, and reacting for 20 to 30 minutes under the condition of 37 DEG C; s4, detecting
fluorescence change in real time, and judging a result through a
fluorescence signal; if a
fluorescence signal is detected, it is indicated that the pepper vein flavivirus exists in the
RNA of the to-be-detected sample, efficient amplification is achieved through a
recombinase-single-chain
binding protein-primer compound, and high sensitivity and specificity of detection are ensured through accurate
nucleic acid recognition and
signal amplification functions; the method is especially suitable for rapid screening and diagnosis of
plant viruses in limited resource environments such as fields.