Digital Point‐of‐Care Testing Device Based on Bioluminescent Pyrophosphate in Real‐Time and Recombinase‐aid Amplification

The breakout of African swine fever virus (ASFV) has severely impacted food safety and public health. Here, a point‐of‐care testing method based on solvent‐responsive magnetic beads suitable for large‐area diagnosis and mutation screening in remote areas is developed to detect it by combining bioluminescent pyrophosphate in real‐time (BART) technology and recombinase‐aid amplification (RAA) technology. The combined testing method includes selection of different deoxyadenosine triphosphate modification structures and DNA polymerases. The method is conducted on plasmids of ASFV at various concentrations. Based on above, the detection device is developed and achieved detection in real‐time. The detection limit is around 10 copies, with no cross‐reaction of nucleic acid samples from other viruses. Then, it is applied on clinical samples and showed good specificity and detection efficiency. Moreover, this method is also compatible with a digital testing method and digital POCT devices based on RAA‐BART device and a mobile phone shell (based on bionic hydrogel) are developed here. Finally, the sensitive of them are tested and showed a comparable detection limit of single copy.