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A CRISPR-Cas12a-based specific enhancer for more sensitive detection of SARS-CoV-2 infection
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.
Affiliation
  • Weiren Huang; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Lei Yu; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Donghua Wen; Department of Laboratory Medicine, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200123, China
  • Dong Wei; Research Laboratory of Clinical Virology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Yangyang Sun; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Huailong Zhao; Jinan Center for Disease Control and Prevention, Jinan, Shandong 250021, China
  • Yu Ye; Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
  • Wei Chen; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Yongqiang Zhu; Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, 201203, China
  • Lijun Wang; Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, 201203, China
  • Li Wang; Jinan Infectious Diseases Hospital of Shandong University, Jinan, Shandong 250021, China
  • Wenjuan Wu; Department of Laboratory Medicine, Shanghai East Hospital, Tongji University School of Medicine
  • Qianqian Zhao; Medical Research & Laboratory Diagnostic Center, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250013, China
  • Yong Xu; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Dayong Gu; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Guohui Nie; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Dongyi Zhu; Department of Laboratory Medicine, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200123, China
  • Zhongliang Guo; Department of Laboratory Medicine, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200123, China
  • Xiaoling Ma; The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230001, China
  • Liman Niu; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Yikun Huang; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Yuchen Liu; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Bo Peng; Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China
  • Renli Zhang; Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China
  • Xiuming Zhang; Shenzhen Sixth People's Hospital (Nanshan Hospital), Huazhong University of Science and Technology Union Shenzhen Hospital, Shenzhen 518052, China
  • Dechang Li; Yuebei Second People's Hospital, Shaoguan, Guangdong 512000, China
  • Yang Liu; Shanghai Institute of Quality Inspection and Technical Research/National Quality Supervision and Inspection Center for Food Products (Shanghai), Shanghai 200233
  • Guoliang Yang; Jinan Center for Disease Control and Prevention, Jinan, Shandong 250021, China
  • Lanzheng Liu; Jinan Center for Disease Control and Prevention, Jinan, Shandong 250021, China
  • Yunying Zhou; Medical Research & Laboratory Diagnostic Center, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250013, China
  • Yunshan Wang; Medical Research & Laboratory Diagnostic Center, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250013, China
  • Tieying Hou; Laboratory of Medicine, Provincial People's Hospital, Guangdong Academy of Medical Sciences Guangzhou, Guangdong 510080, China
  • Qiuping Gao; Tolo Biotechnology Company Limited, Shanghai 200233, China
  • Wujiao Li; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Shuo Chen; Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
  • Xuejiao Hu; Laboratory of Medicine, Provincial People's Hospital, Guangdong Academy of Medical Sciences Guangzhou, Guangdong 510080, China
  • Mei Han; Public Health Medical Centre of Chongqing Municipality, Chongqing 400036, China
  • Huajun Zheng; Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, 201203, China
  • Jianping Wen; The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230001, China
  • Zhiming Cai; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Xinxin Zhang; Research Laboratory of Clinical Virology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Fei Song; Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, International Cancer Center, Shenzhen University
  • Guoping Zhao; CAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Sh
  • Jin Wang; College of Life Sciences, Shanghai Normal University, Shanghai 200234, China
Preprint in En | PREPRINT-MEDRXIV | ID: ppmedrxiv-20119735
Journal article
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ABSTRACT
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. One Sentence SummaryCRISPR-Cas12a-based COVID-19 diagnosis.
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Full text: 1 Collection: 09-preprints Database: PREPRINT-MEDRXIV Type of study: Diagnostic_studies Language: En Year: 2020 Document type: Preprint
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Full text: 1 Collection: 09-preprints Database: PREPRINT-MEDRXIV Type of study: Diagnostic_studies Language: En Year: 2020 Document type: Preprint