Development of a Laboratory-safe and Low-cost Detection Protocol for SARS-CoV-2 of the Coronavirus Disease 2019 (COVID-19)

Won, Joungha; Lee, Solji; Park, Myungsun; Kim, Tai Young; Park, Mingu Gordon; Choi, Byung Yoon; Kim, Dong-Wan; Chang, Hyeshik; Kim, V. Narry; Lee, C. Justin

doi:10.5607/en20009

Cited by 192 publications

(223 citation statements)

References 12 publications

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“…Won et al describe a complete workflow for COVID-19 testing which includes TRIzol extraction and isopropanol precipitation of the RNA from swab samples. The authors found no difference between TRIzol and the approved Qiagen QIAamp Viral RNA Mini Kit in RNA extraction efficiency from Lentivirus-infected HEK293 cells, but was not directly compared using SARS-CoV-2 RNA (Won et al 2020). In our laboratory, isopropanol precipitation of synthesized SARS-CoV-2 viral N gene RNA resulted in almost no loss of RNA, with or without the presence of additional human RNA as a carrier (C Dugast-Darzacq, T Graham, G M Dailey et al, unpublished).…”

Section: Rna Purificationmentioning

confidence: 96%

“…The original CDC protocol approved four commercial master mixes for the RT-PCR test from Quantabio, Promega, and ThermoFisher (Centers for Disease Control and Prevention 2020). However, published RT-PCR protocols have also successfully employed 1-step RT-PCR master mixes from a variety of companies including NEB, Applied Biosciences, Qiagen, Roche, Takara, and others and a growing list of approved alternative commercial reagents can be found at the FDA EUA website (Chandler-Brown et al 2020;Kalikiri et al 2020;Zhao et al 2020;Won et al 2020;Merindol et al;Alcoba-Florez et al 2020;Marzinotto et al;Xu et al 2020;Food and Drug Administration). Many commercial master mixes seem to function well in the detection of SARS-CoV-2, although a detailed side-byside comparison of the numerous commercial reagents is lacking.…”

Section: Rt-pcrmentioning

confidence: 99%

See 1 more Smart Citation

Overcoming the bottleneck to widespread testing: a rapid review of nucleic acid testing approaches for COVID-19 detection

Esbin

Whitney

Chong

et al. 2020

RNA

473

504

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The current COVID-19 pandemic presents a serious public health crisis, and a better understanding of the scope and spread of the virus would be aided by more widespread testing. Nucleicacid based tests currently offer the most sensitive and early detection of COVID-19. However, the "gold standard" test pioneered by the United States Center for Disease Control & Prevention, takes several hours to complete and requires extensive human labor, materials such as RNA extraction kits that could become in short supply and relatively scarce qPCR machines. It is clear that a huge effort needs to be made to scale up current COVID-19 testing by orders of magnitude. There is thus a pressing need to evaluate alternative protocols, reagents, and approaches to allow nucleic-acid testing to continue in the face of these potential shortages. There has been a tremendous explosion in the number of papers written within the first weeks of the pandemic evaluating potential advances, comparable reagents, and alternatives to the "gold-standard" CDC RT-PCR test. Here we present a collection of these recent advances in COVID-19 nucleic acid testing, including both peer-reviewed and preprint articles. Due to the rapid developments during this crisis, we have included as many publications as possible, but many of the cited sources have not yet been peer-reviewed, so we urge researchers to further validate results in their own labs. We hope that this review can urgently consolidate and disseminate information to aid researchers in designing and implementing optimized COVID-19 testing protocols to increase the availability, accuracy, and speed of widespread COVID-19 testing.

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Section: Rna Purificationmentioning

confidence: 96%

Section: Rt-pcrmentioning

confidence: 99%

Overcoming the bottleneck to widespread testing: a rapid review of nucleic acid testing approaches for COVID-19 detection

Esbin

Whitney

Chong

et al. 2020

RNA

473

504

View full text Add to dashboard Cite

show abstract

“…An oropharyngeal swab can be collected but is not essential; if collected, it should be placed in the same container as the nasopharyngeal specimen. An oropharyngeal swab is an acceptable alternative if nasopharyngeal swabs are unavailable [32]. Expectorated sputum should be collected from patients with productive cough; induction of sputum is not recommended.…”

Section: Diagnosismentioning

confidence: 99%

Coronavirus Diseases (COVID-19) Current Status and Future Perspectives: A Narrative Review

Gennaro

Pizzol²,

Marotta

et al. 2020

IJERPH

638

480

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At the end of 2019 a novel virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causing severe acute respiratory syndrome expanded globally from Wuhan, China. In March 2020 the World Health Organization declared the SARS-Cov-2 virus a global pandemic. We performed a narrative review to describe existing literature with regard to Corona Virus Disease 2019 (COVID-19) epidemiology, pathophysiology, diagnosis, management and future perspective. MEDLINE, EMBASE and Scopus databases were searched for relevant articles. Although only when the pandemic ends it will be possible to assess the full health, social and economic impact of this global disaster, this review represents a picture of the current state of the art. In particular, we focus on public health impact, pathophysiology and clinical manifestations, diagnosis, case management, emergency response and preparedness. a natural and zoonotic origin: two scenarios that can plausibly explain the origin of SARS-CoV2 are: (i) natural selection in an animal host before zoonotic transfer; and (ii) natural selection in humans following zoonotic transfer [5,6]. Clinical features and risk factors are highly variable, making the clinical severity range from asymptomatic to fatal [7]. Understanding of COVID-19 is on-going. This review aims to summarize early findings on the epidemiology, clinical features, diagnosis, management, and prevention of COVID-19.

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“…There has also been unprecedented rapid and open sharing of diagnostics and devices through preprint publications, and through informal communication routes among researchers [90,[106][107][108]. For example, agencies in China, Germany, Hong Kong, Japan, Thailand, South Africa and the United States have shared protocols for molecular assays online, including the DNA sequences to detect the virus.…”

Section: Fosh Approaches In Covid-19 Diagnosticsmentioning

confidence: 99%

“…Local manufacturing of health technologies depends on the presence of relevant capacity, infrastructure and regulatory frameworks but is promoted by organisations including the WHO, UNIDO, UNCTAD, UNAIDS, UNICEF and The Global Fund [127]. The implementation of low-cost diagnostic technologies in countries with very limited testing resources could be a game changer in the global control of the COVID-19 pandemic, particularly when combined with protocols for RNA extraction that do not depend on commercial kits ( [90][91][92]119,121], and Table 3), and the generation of reagents that can be distributed and stored at room temperature [128]. For this reason, development of rapid diagnostics for use at the community level was #1 in the WHO's eight immediate research actions agreed as part of their 2019 Novel Coronavirus Global Research and Innovation Forum [129].…”

Section: Fosh Approaches In Covid-19 Diagnosticsmentioning

confidence: 99%

Leveraging open hardware to alleviate the burden of COVID-19 on global health systems

et al. 2020

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With the current rapid spread of COVID-19, global health systems are increasingly overburdened by the sheer number of people that need diagnosis, isolation and treatment. Shortcomings are evident across the board, from staffing, facilities for rapid and reliable testing to availability of hospital beds and key medical-grade equipment. The scale and breadth of the problem calls for an equally substantive response not only from frontline workers such as medical staff and scientists, but from skilled members of the public who have the time, facilities and knowledge to meaningfully contribute to a consolidated global response. Here, we summarise community-driven approaches based on Free and Open Source scientific and medical Hardware (FOSH) as well as personal protective equipment (PPE) currently being developed and deployed to support the global response for COVID-19 prevention, patient treatment and diagnostics.

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Development of a Laboratory-safe and Low-cost Detection Protocol for SARS-CoV-2 of the Coronavirus Disease 2019 (COVID-19)

Cited by 192 publications

References 12 publications

Overcoming the bottleneck to widespread testing: a rapid review of nucleic acid testing approaches for COVID-19 detection

Overcoming the bottleneck to widespread testing: a rapid review of nucleic acid testing approaches for COVID-19 detection

Coronavirus Diseases (COVID-19) Current Status and Future Perspectives: A Narrative Review

Leveraging open hardware to alleviate the burden of COVID-19 on global health systems

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