Streamlined inactivation, amplification, and Cas13-based detection of SARS-CoV-2

Arizti-Sanz, Jon; Freije, Catherine A.; Stanton, Alexandra C.; Petros, Brittany A.; Boehm, Chloe K.; Siddiqui, Sameed M; Shaw, Bennett; Adams, Gordon; Kosoko-Thoroddsen, Tinna Sólveig F.; Kemball, Molly; Uwanibe, Jessica N.; Ajogbasile, Fehintola V.; Eromon, Philomena; Gross, Robin; Wronka, Loni; Caviness, Katie; Hensley, Lisa E.; Bergman, Nicholas H.; MacInnis, Bronwyn; Happi, Christian; Lemieux, Jacob E.; Sabeti, Pardis C.; Myhrvold, Cameron

doi:10.1038/s41467-020-19097-x

Cited by 359 publications

(398 citation statements)

References 40 publications

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“…6). A companion manuscript shows that the improvements in RT-RPA we implemented in eRPA also improve other detection approaches such as SHINE 36 , allowing these assays to become 1-pot, closed-tube, fluorescent readout reactions. eRPA brings us closer to an at-home SARS-CoV-2 test and continued efforts to improve eRPA include: (i) making reactions self-contained and not requiring the addition of reagents before readout, and (ii) making the assay readout quantitative by integration with a lateral flow reader or smartphone app.…”

Section: Discussionmentioning

confidence: 99%

An enhanced isothermal amplification assay for viral detection

et al. 2020

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Rapid, inexpensive, robust diagnostics are essential to control the spread of infectious diseases. Current state of the art diagnostics are highly sensitive and specific, but slow, and require expensive equipment. Here we report the development of a molecular diagnostic test for SARS-CoV-2 based on an enhanced recombinase polymerase amplification (eRPA) reaction. eRPA has a detection limit on patient samples down to 5 viral copies, requires minimal instrumentation, and is highly scalable and inexpensive. eRPA does not cross-react with other common coronaviruses, does not require RNA purification, and takes ~45 min from sample collection to results. eRPA represents a first step toward at-home SARS-CoV-2 detection and can be adapted to future viruses within days of genomic sequence availability.

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Section: Discussionmentioning

confidence: 99%

An enhanced isothermal amplification assay for viral detection

et al. 2020

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“…Similarly, a SHERLOCK assay was validated on >150 patient samples in Thailand having strong concordance with RT-qPCR ( Patchsung et al., 2020 ). Many more publications soon followed ( Ackerman et al., 2020 ; Ali et al., 2020 ; Arizti-Sanz et al., 2020 ; Chen et al., 2020 ; Fozouni et al., 2021 ; Guo et al., 2020 ; Hou et al., 2020 ; Huang et al., 2020 ; Joung et al., 2020 ; Ning et al., 2020 ; Ramachandran et al., 2020 ; Wang et al., 2020b ) with methods summarized in Table 1 .…”

Section: Crispr-based Detection Of Virusesmentioning

confidence: 99%

Detect and destroy: CRISPR-based technologies for the response against viruses

Freije

Sabeti

2021

Cell Host & Microbe

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Despite numerous viral outbreaks in the last decade, including a devastating global pandemic, diagnostic and therapeutic technologies remain severely lacking. CRISPR-Cas systems have the potential to address these critical needs in the response against infectious disease. Initially discovered as the bacterial adaptive immune system, these systems provide a unique opportunity to create programmable, sequence-specific technologies for detection of viral nucleic acids and inhibition of viral replication. This review summarizes how CRISPR-Cas systems—in particular the recently discovered DNA-targeting Cas12 and RNA-targeting Cas13, both possessing a unique trans -cleavage activity—are being harnessed for viral diagnostics and therapies. We further highlight the numerous technologies whose development has accelerated in response to the COVID-19 pandemic.

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“…No thermo-cycle help reduce the potential cross-sample aerosol contamination within the equipment and detection facilities. In Cas13-based detection system optimized by Arizti-Sanz et al [ 49 ], it was validated with 90% sensitivity and 100% specificity with a turnaround time of 50 minutes. It is more exciting that CRISPR-Cas12a assay could bring about an ultrasensitive, instrument-free, and visual detection (for few copies) within as soon as 20 minutes [ 50 ].…”

Section: Nucleic Acid Detection Of Sars-cov-2mentioning

confidence: 99%

Molecular detection of SARS-CoV-2 being challenged by virus variation and asymptomatic infection

Jiang

et al. 2021

Journal of Pharmaceutical Analysis

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Coronavirus disease 2019 (COVID-19) has been a pandemic for more than a year. With the expanding second wave of pandemic in winter, the continuous evolution of SARS-CoV-2 has brought new issues, including the significance of virus mutations in infection and the detection of asymptomatic infection. In this review, we firstly introduced several major SARS-CoV-2 mutations since the COVID-19 outbreak and then mentioned the widely used molecular detection techniques to diagnose COVID-19, primarily focusing on their strengths and limitations. We further discussed the effects of viral genetic variation and asymptomatic infection on the molecular detection of SARS-CoV-2 infection. The review finally summarized useful insights into the molecular diagnosis of COVID-19 under the special situation challenging by virus mutation and asymptomatic infection.

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Streamlined inactivation, amplification, and Cas13-based detection of SARS-CoV-2

Cited by 359 publications

References 40 publications

An enhanced isothermal amplification assay for viral detection

An enhanced isothermal amplification assay for viral detection

Detect and destroy: CRISPR-based technologies for the response against viruses

Molecular detection of SARS-CoV-2 being challenged by virus variation and asymptomatic infection

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