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All-in-One Dual CRISPR-Cas12a (AIOD-CRISPR) Assay: A Case for Rapid, Ultrasensitive and Visual Detection of Novel Coronavirus SARS-CoV-2 and HIV virus

View ORCID ProfileXiong Ding, Kun Yin, Ziyue Li, Changchun Liu
doi: https://doi.org/10.1101/2020.03.19.998724
Xiong Ding
Department of Biomedical Engineering, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT, 06030, United States
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  • ORCID record for Xiong Ding
Kun Yin
Department of Biomedical Engineering, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT, 06030, United States
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Ziyue Li
Department of Biomedical Engineering, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT, 06030, United States
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Changchun Liu
Department of Biomedical Engineering, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT, 06030, United States
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  • For correspondence: chaliu@uchc.edu
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Abstract

A recent outbreak of novel coronavirus (SARS-CoV-2), the causative agent of COVID-19, has spread rapidly all over the world. Human immunodeficiency virus (HIV) is another deadly virus and causes acquired immunodeficiency syndrome (AIDS). Rapid and early detection of these viruses will facilitate early intervention and reduce disease transmission risk. Here, we present an All-In-One Dual CRISPR-Cas12a (termed “AIOD-CRISPR”) assay method for simple, rapid, ultrasensitive, one-pot, and visual detection of coronavirus SARS-CoV-2 and HIV virus. In our AIOD CRISPR assay, a pair of crRNAs was introduced to initiate dual CRISPR-Cas12a detection and improve detection sensitivity. The AIOD-CRISPR assay system was successfully utilized to detect nucleic acids (DNA and RNA) of SARS-CoV-2 and HIV with a sensitivity of few copies. Also, it was evaluated by detecting HIV-1 RNA extracted from human plasma samples, achieving a comparable sensitivity with real-time RT-PCR method. Thus, our method has a great potential for developing next-generation point-of-care molecular diagnostics.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted March 21, 2020.
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All-in-One Dual CRISPR-Cas12a (AIOD-CRISPR) Assay: A Case for Rapid, Ultrasensitive and Visual Detection of Novel Coronavirus SARS-CoV-2 and HIV virus
Xiong Ding, Kun Yin, Ziyue Li, Changchun Liu
bioRxiv 2020.03.19.998724; doi: https://doi.org/10.1101/2020.03.19.998724
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All-in-One Dual CRISPR-Cas12a (AIOD-CRISPR) Assay: A Case for Rapid, Ultrasensitive and Visual Detection of Novel Coronavirus SARS-CoV-2 and HIV virus
Xiong Ding, Kun Yin, Ziyue Li, Changchun Liu
bioRxiv 2020.03.19.998724; doi: https://doi.org/10.1101/2020.03.19.998724

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