Ultrafast visual nucleic acid detection with CRISPR/Cas12a and rapid PCR in single capillary

Abstract

Rapid and sensitive nucleic acid detection is critical for point-of-care testing (POCT). Here, we proposed an ultrasensitive visual nucleic acid detection method that could be accomplished in <10 min. By integrating rapid PCR amplification and Cas12a cleavage as one-pot reaction in capillary, the operation was greatly simplified and amplicon contamination was totally avoided. Rapid PCR amplification was performed in portable thermo cups and Cas12a cleavage reaction was triggered with body-heat. Fluorescent results could be easily identified by naked eye with the help of a mini UV-touch. Results indicated that the proposed method possesses detection sensitivity at single molecule level, with high robustness, specificity and reliability. Thus, we have created a field-deployable nucleic acid detection platform with high performance and minimal equipment. It has broad application prospects for POCT in fields ranging from food safety determination to clinical diagnosis.