Establishment of RPA/CRISPR-Cas12a based rapid detection for Verticillium alfalfae
Verticillium alfalfae is an important quarantine pathogen,which can cause serious damage to more than a dozen crops.The sequence characterized amplified region(SCAR)marker of V.alfalfae was chosen as the target gene,and a detection method combining RPA isothermal amplification technique and CRISPR-Cas12a system was established for specific detection of V.alfalfae.Under the optimized experimental conditions,10.6 pg genomic DNA can be detected by using CRISPR-Cas12a based fluorescence and lateral flow strip method after the genomic DNA was amplified at 39℃ for 30 minutes.The spiked recovery experiment indicated that the established RPA/CRISPR-Cas12a can be used for the rapid detection of V.alfalfae in plant tissues.
万方数据
维普
