CRISPR/Cas12a-核酸等温扩增技术在食源性致病菌检测中的研究进展OA

The CRISPR/Cas12a gene editing system can efficiently recognize and cleave exogenous nucleic acids,thus driving the innovation and development of nucleic acid-based detection technologies for foodborne pathogens.This review summarizes the research progress on integrating CRISPR/Cas 12a-based detection with various isothermal amplification techniques,including loop-mediated isothermal amplification(LAMP),recombinase polymerase amplification(RPA),recombinase-mediated amplification(RAA),rolling circle amplification(RCA),hybridization chain reaction(HCR),strand displacement amplification(SDA),and exponential amplification reaction(EXPAR),for the detection of foodborne pathogens.The combination of CRISPR/Cas12a with various isothermal amplification techniques enables highly specific and sensitive visual detection of foodborne pathogens,offering advantages such as simple operation,rapidity,and no requirement for complex instrumentation,while still facing limitations including high reagent costs,narrow temperature control range,relatively complex procedures,stringent primer design requirements,and low standardization.Future directions for the integration of CRISPR/Cas12a with isothermal amplification techniques focus on improving detection throughput and accuracy,developing automated and integrated platforms,and leveraging smartphone-based and artificial intelligence-based systems,thereby enabling"sample-in,result-out"one-pot on-site detection and providing technical support for more efficient,sensitive,simple,and rapid detection of foodborne pathogens.