A CRISPR-Cas12a-based specific enhancer for more sensitive detection of SARS-CoV-2 infection
By
Weiren Huang,
Lei Yu,
Donghua Wen,
Dong Wei,
Yangyang Sun,
Huailong Zhao,
Yu Ye,
Wei Chen,
Yongqiang Zhu,
Lijun Wang,
Li Wang,
Wenjuan Wu,
Qianqian Zhao,
Yong Xu,
Dayong Gu,
Guohui Nie,
Dongyi Zhu,
Zhongliang Guo,
Xiaoling Ma,
Liman Niu,
Yikun Huang,
Yuchen Liu,
Bo Peng,
Renli Zhang,
Xiuming Zhang,
Dechang Li,
Yang Liu,
Guoliang Yang,
Lanzheng Liu,
Yunying Zhou,
Yunshan Wang,
Tieying Hou,
Qiuping Gao,
Wujiao Li,
Shuo Chen,
Xuejiao Hu,
Mei Han,
Huajun Zheng,
Jianping Wen,
Zhiming Cai,
Xinxin Zhang,
Fei Song,
Guoping Zhao,
Jin Wang
Posted 05 Jun 2020
medRxiv DOI: 10.1101/2020.06.02.20119735
High Ct-values falling in the grey zone are frequently encountered in SARS-CoV-2 detection by real-time reverse transcription PCR (rRT-PCR) and have brought urgent challenges in diagnosis of samples with low viral load. Based on the single-stranded DNA reporter trans-cleavage activity by Cas12a upon target DNA recognition, we create a Specific Enhancer for detection of PCR-amplified Nucleic Acids (SENA) to confirm SARS-CoV-2 detection through specifically targeting its rRT-PCR amplicons. SENA is highly sensitive, with its limit of detection being at least 2 copies/reaction lower than that of the corresponding rRT-PCR, and highly specific, which identifies both false-negative and false-positive cases in clinic applications. SENA provides effective confirmation for nucleic acid amplification-based molecular diagnosis, and may immediately eliminate the uncertainty problems of rRT-PCR in SARS-CoV-2 clinic detection.
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