A signal on–off strategy based on the digestion of DNA cubes assisted by the CRISPR–Cas12a system for ultrasensitive HBV detection in solid-state nanopores
Jiahai Wang, Lisiteng Luo, Yunhui Li, Lanfang Chen, Cenlin Gui, Jianji Zhu, Huizhen Li, Wenlong Wang, Daqi Chen
Abstract
hepatitis B virus (HBV) target-triggered cleavage of the component elements of the DNA cube with the assistance of the CRISPR-Cas12a technology, which caused a great drop in the translocation rate. The elements to cleave were optimized, and the sensor performance was tested in different protein stabilizer-rich buffers and human serum. Coupling with the polymerase chain reaction (PCR) pre-amplification technology, HBV-positive or -negative classification was achieved with the detection limit reaching 5 aM. It is worth noting that in our method, all reaction buffers were directly used without further optimization, which is of great help for the practical application of solid-state nanopores.