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Development and Evaluation of A CRISPR-based Diagnostic For 2019-novel Coronavirus

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

Posted: February 23, 2020
Server: medRxiv
DOI: 10.1101/2020.02.22.20025460

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 for 2019-nCoV.

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 the novel virus. The diagnostic performance of CRISPR-nCoV was assessed and also 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 withSARS-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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