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.
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