Photoactivatable CRISPR/Cas9 lateral flow strip platform for one-pot rapid detection of squamous cell carcinoma antigen DNA in blood

Abstract

Cervical cancer, one of the major threats to women's health, has been gradually decreasing in age of onset and shows a younger trend in recent years. Squamous cell carcinoma antigen (SCCA), as the preferred serum biomarker of cervical cancer, can be effectively screened and detected by quantitative polymerase chain reaction (qPCR). However, the operation process of qPCR is time-consuming and needs to rely on complicated experimental procedures. In this study, we creatively developed a UV light-controlled CRISPR/Cas9 lateral flow strip system for one-pot rapid detection of SCCA DNA in 34 clinical peripheral blood samples. Compared with the commercial SCCA ELISA kit, the photoactivatable detection system not only simplified the whole process without DNA extraction, but also completed the true one-pot rapid detection in 30 minutes, effectively preventing the occurrence of off-target effect and avoiding the risk of sample contamination during the process. In addition, we have successfully solved the incompatibility between recombinase-aided amplification (RAA) and CRISPR system, and increased the minimum detection limit to 10 fg/μL. The detection accuracy has reached an amazing 94.12 %, which is much higher than 79.41 % of the SCCA ELISA kit. Our study indicates that this photoactivatable CRISPR/Cas9 lateral flow strip system has potential application prospects for the early diagnosis of cervical cancer, and further extended to the detection of other related diseases.