《MedRxiv,2月23日,Development and Evaluation of A CRISPR-based Diagnostic For 2019-novel Coronavirus》

  • 来源专题:COVID-19科研动态监测
  • 编译者: xuwenwhlib
  • 发布时间:2020-02-24
  • Development and Evaluation of A CRISPR-based Diagnostic For 2019-novel Coronavirus

    Tieying Hou, Weiqi Zeng, Minling Yang, Wenjing Chen, Lili Ren, Jingwen Ai, Ji Wu, Yalong Liao, Xuejing Gou, Yongjun Li, Xiaorui Wang, Hang Su, Jianwei Wang, Bing Gu, Teng Xu

    doi: https://doi.org/10.1101/2020.02.22.20025460

    Abstract

    Background: The recent outbreak of infections by the 2019 novel coronavirus (2019-nCoV), the third zoonotic CoV has raised great public health concern. The demand for rapid and accurate diagnosis of this novel pathogen brought significant clinical and technological challenges. Currently, metagenomic next-generation sequencing (mNGS) and reverse-transcription PCR (RT-PCR) are the most widely used molecular diagnostics. Methods: 2019-nCoV infections were confirmed in 52 specimens by mNGS. Genomic information was analyzed and used for the design and development of an isothermal, CRISPR-based diagnostic for this novel virus. The diagnostic performance of CRISPR-nCoV was assessed and compared across three technology platforms (mNGS, RT-PCR and CRISPR). Results: 2019-nCoVs sequenced in our study were conserved with the Wuhan strain, and shared certain genetic similarity with SARS-CoV. A high degree of variation in the level of viral RNA was observed in clinical specimens. CRISPR-nCoV demonstrated a near single-copy sensitivity and great clinical sensitivity with a shorter turn-around time than RT-PCR. Conclusion: CRISPR-nCoV presents as a promising diagnostic option for the emerging pathogen.

    *注,本文为预印本论文手稿,是未经同行评审的初步报告,其观点仅供科研同行交流,并不是结论性内容,请使用者谨慎使用.

  • 原文来源:https://www.medrxiv.org/content/10.1101/2020.02.22.20025460v1
相关报告
  • 《MedRxiv,3月10日,Rapid Detection of 2019 Novel Coronavirus SARS-CoV-2 Using a CRISPR-based DETECTR Lateral Flow Assay》

    • 来源专题:COVID-19科研动态监测
    • 编译者:zhangmin
    • 发布时间:2020-03-11
    • Rapid Detection of 2019 Novel Coronavirus SARS-CoV-2 Using a CRISPR-based DETECTR Lateral Flow Assay James P Broughton, Xianding Deng, Guixia Yu, Clare L Fasching, Jasmeet Singh, Jessica Streithorst, Andrea Granados, Alicia Sotomayor-Gonzalez, Kelsey Zorn, Allan Gopez, Elaine Hsu, Wei Gu, Steven Miller, Chao-Yang Pan, Hugo Guevara, Debra Wadford, Janice Chen, Charles Y Chiu doi: https://doi.org/10.1101/2020.03.06.20032334 Abstract An outbreak of novel betacoronavirus, SARS-CoV-2 (formerly named 2019-nCoV), began in Wuhan, China in December 2019 and the COVID-19 disease associated with infection has since spread rapidly to multiple countries. Here we report the development of SARS-CoV-2 DETECTR, a rapid (~30 min), low-cost, and accurate CRISPR-Cas12 based lateral flow assay for detection of SARS-CoV-2. We validated this method using contrived reference samples and clinical samples from infected US patients and demonstrated comparable performance to the US CDC SARS-CoV-2 real-time RT-PCR assay. *注,本文为预印本论文手稿,是未经同行评审的初步报告,其观点仅供科研同行交流,并不是结论性内容,请使用者谨慎使用.
  • 《MedRxiv,2月18日,Clinical diagnosis of 8274 samples with 2019-novel coronavirus in Wuhan》

    • 来源专题:COVID-19科研动态监测
    • 编译者:xuwenwhlib
    • 发布时间:2020-02-19
    • Clinical diagnosis of 8274 samples with 2019-novel coronavirus in Wuhan Ming Wang, Qing Wu, Wanzhou Xu, Bin Qiao, Jingwei Wang, Hongyun Zheng, Shupeng Jiang, Junchi Mei, Zegang Wu, Yayun Deng, Fangyuan Zhou, Wei Wu, Yan Zhang, Zhihua Lv, Jingtao Huang, Xiaoqian Guo, Lina Feng, Zunen Xia, Di Li, Zhiliang Xu, Tiangang Liu, Pingan Zhang, Yongqing Tong, Yan Li doi: https://doi.org/10.1101/2020.02.12.20022327 Abstract Background: 2019-Novel coronavirus (2019-nCoV) outbreaks create challenges for hospital laboratories because thousands of samples must be evaluated each day. Sample types, interpretation methods, and corresponding laboratory standards must be established. The possibility of other infections should be assessed to provide a basis for clinical classification, isolation, and treatment. Accordingly, in the present study, we evaluated the testing methods for 2019-nCoV and co-infections. Methods: We used a fluorescence-based quantitative PCR kit urgently distributed by the Chinese CDC to detect 8274 close contacts in the Wuhan region against two loci on the 2019-nCoV genome. We also analyzed 613 patients with fever who underwent multiple tests for 13 respiratory pathogens; 316 subjects were also tested for 2019-nCoV. Findings: Among the 8274 subjects, 2745 (33.2%) had 2019-nCoV infection; 5277 (63.8%) subjects showed negative results in the 2019-nCoV nucleic acid test (non-019-nCoV); and 252 cases (3.0%) because only one target was positive, the diagnosis was not definitive. Sixteen patients who originally had only one positive target were re-examined a few days later; 14 patients (87.5%) were finally defined as 2019-nCoV-positive, and 2 (12.5%) were finally defined as negative. The positive rates of nCoV-NP and nCovORF1ab were 34.7% and 34.7%, respectively. nCoV-NP-positive only and nCovORF1ab-positive cases accounted for 1.5% and 1.5%, respectively. In the 316 patients with multiple respiratory pathogens, 104 were positive for 2019-nCov and 6/104 had co-infection with coronavirus (3/104), influenza A virus (2/104), rhinovirus (2/104), and influenza A H3N2 (1/104); the remaining 212 patients had influenza A virus (11/202), influenza A H3N2 (11/202), rhinovirus (10/202), respiratory syncytial virus (7/202), influenza B virus (6/202), metapneumovirus (4/202), and coronavirus (2/202). Interpretation: Clinical testing methods for 2019-nCoV require improvement. Importantly, 5.8% of 2019-nCoV infected and 18.4% of non-2019-nCoV-infected patients had other pathogen infections. It is important to treat combined infections and perform rapid screening to avoid cross-contamination of patients. A test that quickly and simultaneously screens as many pathogens as possible is needed. *注,本文为预印本论文手稿,是未经同行评审的初步报告,其观点仅供科研同行交流,并不是结论性内容,请使用者谨慎使用.