Method for visually detecting maize chlorotic mottle virus based on RAA-CRISPR/Cas12a technology

• Main Authors: MA WENDI, SUN XI, FAN ZAIFENG, TIAN YIYING, JIAO ZHIYUAN, DUAN XUEYAN, ZHOU TAO
• Format: Patent
• Language: Chinese; English
• Published: 08.08.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a method and a kit for visually detecting a maize chlorotic mottle virus based on a RAA-CRISPR/Cas12a technology and application of the method and the kit. The kit comprises an RAA primer pair, an ssDNA probe, crRNA and Cas12a protein, the RAA primer pair is composed of an MCMV-F1 primer and an MCMV-R3 primer, the MCMV-F1 primer is a single-stranded DNA molecule as shown in SEQ ID No.1 in a sequence table, the MCMV-R3 primer is a single-stranded DNA molecule as shown in SEQ ID No.4 in the sequence table, the ssDNA probe is a single-stranded DNA molecule as shown in SEQ ID No.13 in the sequence table, and the crRNA is an RNA molecule as shown in SEQ ID No.12 in the sequence table. The kit is high in detection sensitivity, high in amplification speed, short in reaction time, efficient, simple and convenient, does not need complex instruments and equipment, has macroscopic detection results, and is suitable for on-site rapid detection of the maize chlorotic mottle virus. 本发明公开了基于RAA-CRISP