SARS-CoV-2 On-the-Spot Virus Detection Directly from Patients

Ben-Assa, Nadav; Naddaf, Rawi; Gefen, Tal; Capucha, Tal; Hajjo, Haitham; Mandelbaum, Noa; Elbaum, Lilach; Rogov, Peter; Daniel, King A.; Kaplan, Shai; Rotem, Assaf; Chowers, Michal; Szwarcwort-Cohen, Moran; Geva‐Zatorsky, Naama

doi:10.1101/2020.04.22.20072389

Cited by 28 publications

(32 citation statements)

References 10 publications

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“…Colorimetric RT-LAMP on saliva has broad potential to increase COVID-19 screening speed and capacity, as demonstrated by our adaptation of this assay to a plate-based format. Similar approaches by others have now been validated on clinical samples, deployed for disease surveillance, and adapted for home testing and point-of-care use 27,28 .…”

Section: Discussionmentioning

confidence: 99%

Rapid and extraction-free detection of SARS-CoV-2 from saliva with colorimetric LAMP

Lalli¹,

Langmade²,

Chen³

et al. 2020

Preprint

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Rapid, reliable, and widespread testing is required to curtail the ongoing COVID-19 pandemic. Current gold standard diagnostic assays are hampered by supply shortages in critical reagents including nasal swabs, RNA extraction kits, personal protective equipment (PPE), instrumentation, and labor. Here we present an approach to overcome these challenges with the development of a rapid colorimetric assay using reverse-transcription loop-mediated isothermal amplification (RT-LAMP) optimized on human saliva samples without an RNA purification step. We describe our optimizations of the LAMP reaction and saliva pre-treatment protocols that enabled rapid and sensitive detection of < 10 2 viral genomes per reaction in contrived saliva controls. We also observed high performance of this assay on a limited number of clinical saliva samples. While thorough validation on additional clinical samples will be needed before such an assay can be widely used, these preliminary results demonstrate a promising approach to overcome the current bottlenecks limiting widespread testing.

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Section: Discussionmentioning

confidence: 99%

Rapid and extraction-free detection of SARS-CoV-2 from saliva with colorimetric LAMP

Lalli¹,

Langmade²,

Chen³

et al. 2020

Preprint

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“…Other isothermal methods have been proposed for SARS-CoV-2 detection. Most of them are based on the RT-LAMP technology with a colourimetric or turbidimetric readout [7][8][9][10]. These assays could potentially suffer from false positive results generated from nonspecific primer binding or primer dimers [11].…”

Section: Discussionmentioning

confidence: 99%

“…For the second stage, using the 'all pool' results shown in Figure 4c, we calculated that the NGS sequencing alone has a LoD-95 of 80.3 copies (CI = [37. 7,197]) per 20 µl reaction. By combining the Stage 1 fluorescence readout and Stage 2 'negative pool' results, the overall INSIGHT technology LoD-95 can be further improved to 49.9 (CI = [21.2, 125]).…”

Section: Insight Performance Evaluation and Comparison With Rt-qpcrmentioning

confidence: 99%

“…This would allow for decentralised and frequent testing of a large proportion of the population, even in countries with limited medical resources. Numerous groups are working on near-patient tests [7][8][9][10][11][12][13] aimed at improving testing capacity and ultimately achieving regular populational scale testing. However, it is difficult to control for patient operational error and it is also challenging for centralised data collection.…”

Section: Introductionmentioning

confidence: 99%

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INSIGHT: a population scale COVID-19 testing strategy combining point-of-care diagnosis with centralised high-throughput sequencing

Suo

Brown

et al. 2020

Preprint

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We present here INSIGHT (Isothermal NASBA-Sequencing based hIGH-througput Test): a two-stage COVID-19 testing strategy, using a combination of an isothermal NASBA reaction and next generation sequencing. From commercially acquired human saliva with spiked-in viral RNA as input, the first stage employs isothermal amplification of viral RNA to give a rapid result in one to two hours, using either fluorescence detection or a dipstick readout, whilst simultaneously incorporating sample-specific barcodes into the amplification product. In the first stage, fluorescent viral RNA detection can be consistently achieved at 10-100 copies per 20 µl reaction. The second stage pools post-amplification barcoded products from multiple samples for scalable sequencing that could be centralised, to further improve the accuracy of the test in a massively parallel way. Our two-stage testing strategy is suitable for further development into a home-based or point-of-care assay, and is potentially scalable to population level. Affiliations:* Molecular beacons were synthesised in the ATDBio laboratory using a K&A H-8 SE DNA Synthesiser, and purified by reversed-phase HPLC.

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“…The advantage of this method lies in its simplicity which excludes RNA extraction and purification and can be performed in a single step. [82] Moreover, this test showed potential to be used as a surveillance test and provides comparable sensitivity to PCR for detection of COVID-19.…”

Section: Nucleic Acid Testmentioning

confidence: 99%

A Concise Review of Baseline Facts of SARS‐CoV‐2 for Interdisciplinary Research

2020

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The COVID‐19 pandemic caused by SARS‐CoV‐2 is a global health emergency. Investigation covering various aspects of diagnosis, treatment, and vaccine development for COVID‐19 are being attempted globally. Common goal of reducing loss of life requires accelerated and integrated effort from researchers of different fields i. e. chemistry, biology, clinical/medical science etc. Integrated yet timely effort requires the understanding of interdisciplinary key concepts as well as knowledge of major challenges, present status and strategies. Hence, in this article, we present an overview of baseline/key facts of SARS‐CoV‐2 mingled with current status and strategies in a simple, straight forward and coherent manner. In particular, the review attempts to integrate and arrange the key facts at the interface of chemistry and biology. Overview of epidemiology of SARS‐CoV‐2 virus infection, structural characteristics of virus, detection methods and therapeutic agents based on antibodies, therapeutic agents based on traditional drug molecule, drug development based on specific catalytic enzyme inhibitors, vaccine design approaches have been provided. Since enzyme site and catalytic action is the key for biological process, they have been lucidly explained at molecular level. It is believed that accredited information amalgamated in this report will provide a strong rational and lucid groundwork for the coherent interdisciplinary research to countermeasure covid‐19 crisis.

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SARS-CoV-2 On-the-Spot Virus Detection Directly from Patients

Cited by 28 publications

References 10 publications

Rapid and extraction-free detection of SARS-CoV-2 from saliva with colorimetric LAMP

Rapid and extraction-free detection of SARS-CoV-2 from saliva with colorimetric LAMP

INSIGHT: a population scale COVID-19 testing strategy combining point-of-care diagnosis with centralised high-throughput sequencing

A Concise Review of Baseline Facts of SARS‐CoV‐2 for Interdisciplinary Research

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