AUTHOR=Wang Xinge , Chen Yangcan , Cheng Xuejia , Wang Si-Qi , Hu Yanping , Feng Yingmei , Jin Ronghua , Zhou Kangping , Liu Ti , Wang Jianxing , Pan Kai , Liu Bing , Xiang Jie , Wang Yanping , Zhou Qi , Zhang Ying , Pan Weiye , Li Wei 

TITLE=CDetection.v2: One-pot assay for the detection of SARS-CoV-2

JOURNAL=Frontiers in Microbiology

VOLUME=Volume 14 - 2023

YEAR=2023

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2023.1158163

DOI=10.3389/fmicb.2023.1158163

ISSN=1664-302X

ABSTRACT=The ongoing 2019 Coronavirus Disease pandemic (COVID-19), caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) and its variants, is a global public health threat. Early diagnosis and identification of SARS-CoV-2 and its variants plays a critical role in COVID-19 prevention and control. Currently, the most widely used technique to detect SARS-CoV-2 is quantitative reverse transcription PCR (RT-qPCR), which takes nearly 1 hour and should be performed by experienced personnel to ensure the accuracy of results. Therefore, the development of a nucleic acid detection kit with higher sensitivity, faster detection and greater accuracy is important. Here, we developed a Cas12b-assisted one-pot detection platform (CDetection.v2) that allows rapid detection of SARS-CoV-2 in 30 minutes. This platform was able to detect up to 5 × 103 copies/ml of SARS-CoV-2 without cross-reactivity with other viruses. Moreover, the sensitivity of this CRISPR system was comparable to that of RT-qPCR when tested on 120 clinical samples. The CDetection.v2 provides a novel one-pot detection approach based on the integration of RT-RAA and CRISPR/Cas12b for detecting SARS-CoV-2 and screening of large-scale clinical samples, offering a more efficient strategy for detecting various types of viruses.