posted on 2023-11-16, 16:36authored byFukai Wang, Xiaowei Ma, Jing Ye, Chenzhi Shi, Yun Chen, Zhicai Yu, Tianming Li, Donglei Yang, Min Li, Pengfei Wang
Detection of viral infections (e.g., SARS-CoV-2) with
high precision
is critical to disease control and treatment. There is an urgent need
to develop point-of-care detection methods to complement the gold
standard laboratory-based PCR assay with comparable sensitivity and
specificity. Herein, we developed a method termed mCAD to achieve
ultraspecific point-of-care detection of SARS-CoV-2 RNA while maintaining
high sensitivity by programming multiplex rolling circle amplification
and toehold-mediated strand displacement reactions. RCA offers sufficient
amplification of RNA targets for subsequent detection. Most importantly,
a multilayer of detection specificity is implemented into mCAD via
sequence-specific hybridization of nucleic acids across serial steps
of this protocol to fully eliminate potential false-positive detections.
Using mCAD, we demonstrated a highly specific, sensitive, and convenient
visual detection of SARS-CoV-2 RNA from both synthetic and clinical
samples, exhibiting performance comparable to qPCR. We envision that
mCAD will find its broad applications in clinical prospects for nucleic
acid detections readily beyond SARS-CoV-2 RNA.