A flexible RPA-Cas12a strategy for the rapid detection of Varicella zoster virus

• Combining RPA and collateral activity of LbCas12, ultrasensitive diagnosis of VZV is achieved within 40 min. • We developed a one-pot LbCas12a-RPA-LFT detection system by utilizing RPA, LbCas12a and lateral-flow-test (LFT). • Clinical samples including peripheral blood can be effectively identified by one-pot LbCas12a-RPA-LFT within 25 min. • The one-pot LbCas12a-RPA-LFT system may shed light on the development of new POCT platforms for early diagnosis of VZV. Varicella-Zoster Virus (VZV) only infects human beings, largely impairs patients’ living quality and causes health-threatening diseases because of various complications. Timely and accurate diagnosis combined with anti-viral treatment is of great significance for patients infected with VZV. However, on account of the characteristics of low viral load in VZV infection, traditional methods, qPCR and clinical technologies, usually provide false negative results, especially detecting peripheral blood samples. In this study, by combining recombinase polymerase amplification (RPA) and collateral activity of LbCas12a, we established a sequential two-step LbCas12a-RPA system with a detection limit of 3 copies/μL. Comparing to qPCR, LbCas12a-RPA exhibited higher sensitivity, specificity, and accuracy, can accomplish the test within 40 min in laboratory. To achieve point-of-care testing (POCT) diagnosis of VZV, we further utilized lateral flow test (LFT) to develop a visible one-pot system LbCas12a-RPA-LFT showing excellent performance for analyzing VZV clinical samples including saliva and peripheral samples within 25 min. Our one-pot system provides a fast and accurate POCT method for VZV detection.