An ultrasensitive and contamination-free on-site nucleic acid detection platform for Listeria monocytogenes based on the CRISPR-Cas12a system combined with recombinase polymerase amplification

CE–RAA–CRISPR Assay: A Rapid and Sensitive Method for Detecting Vibrio parahaemolyticus in Seafood

“…In the experiment performed by Yin et al [ 32 ], this concentration was less effective for enhancing the fluorescence signal, presumably because the one-pot detection method introduces more amplification products. As reported by several authors [ 20 , 30 , 33 ], the optimal Cas/crRNA ratio varies among targets or detection systems. Li et al [ 25 ] demonstrated the importance of the Cas/crRNA stoichiometry, as the presence of excess crRNA reduces the fluorescence signal.…”

“…The CRISPR/Cas12a system further enhances nucleic acid detection. Although traditional methods based on CRISPR systems, such as HOLMES [ 28 ] and DETECTR [ 29 ], have been successfully applied in nucleic acid detection, they usually require the transfer of recombinase polymerase amplification or PCR amplification products, which increases operational complexity and the risk of sample cross-contamination [ 20 , 30 ]. In our study, CE–RAA–CRISPR, a convenient and highly sensitive method for the rapid one-pot detection of V. parahaemolyticus , was established.…”

“…Thus, through the fluorophore and quencher labeling of the ssDNA, the content of the target gene can be converted into a fluorescent signal [ 17 , 18 , 19 ]. The CRISPR system has been combined with the RAA assay to detect various foodborne pathogens, including Listeria monocytogenes [ 20 ], Salmonella [ 21 ], and spoilage bacteria [ 22 ].…”

Ultra-Sensitive and Rapid Detection of Pathogenic Yersinia enterocolitica Based on the CRISPR/Cas12a Nucleic Acid Identification Platform

“…While the CRISPR/Cas12a detection platform has been applied to many foodborne pathogenic bacteria ( Escherichia coli , Listeria monocytogenes , Staphylococcus aureus , etc. ), an equivalent procedure has not been reported for Y. enterocolitica [ 35 , 36 , 37 ].…”

A Novel Photoelectrochemical Aptamer Sensor Based on CdTe Quantum Dots Enhancement and Exonuclease I-Assisted Signal Amplification for Listeria monocytogenes Detection

“…It is widely found in milk; meat; aquatic products; and frozen, and cold storage, and ready-to-eat foods [ 4 ]. Listeria monocytogenes can cause meningitis and sepsis, which have high hospitalization rates and mortality [ 5 , 6 , 7 ]. Various countries have formulated relevant limit standards.…”