RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2

Huang, Wei E.; Lim, Boon Huat; Hsu, Chia Chen; Xiong, Dan; Wu, Wei; Yu, Yejiong; Jia, Haidong; Wang, Yun; Zeng, Yida; Ji, Mengmeng; Chang, Hong; Zhang, Xiuming; Wang, Hui; Cui, Zhanfeng

doi:10.1111/1751-7915.13586

Cited by 459 publications

(456 citation statements)

References 26 publications

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“…For e cient SARS-CoV-2 screening of every person in our hospital, i.e. all patients and the whole staff, as well as for realtime results we decided to use a point-of-cate device based on LAMP-technology as recommended by others [6,12].…”

Section: Discussionmentioning

confidence: 99%

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VariplexTM test system fails to reliably detect SARS-CoV-2 directly from respiratory samples without RNA extraction

Eckel

Küsters

Drossel

et al. 2020

Preprint

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Diagnosis of COVID is performed by PCR methods, but their capacitiy is limited by the requirement of high-level facilities and instruments. The loop-mediated isothermal amplification (LAMP) method has been utilized for the detection of isolated virus-specific RNA. Preliminary data suggest the possibility of isothermal amplification directly from respiratory samples without RNA extraction.All patients admitted to our hospital were screened for SARS-CoV-2 by routine. Respiratory samples were tested by variplex system based on LAMP method directly without RNA extraction and by PCR. Confirmation or exclusion of clinical COVID diagnosis was based on history, examination and findings. Primary endpoint was the false negative rate of variplex test compared to PCR.A total of 153 patients were included in the study. Median age was 79 years and male/female ratio was 45%/55%. Prevalence of clinical COVID diagnosis was 61.4%. Variplex test and PCR assay was positive in 15.0% and 45.5%, respectively. In 109 patients variplex test and PCR assay were performed simultaneously. False negative rate of variplex test compared to PCR was 83.0%.The potential of LAMP-technology using isolated RNA has been demonstrated impressively by others and excellent sensitivity and specivity of detecting SARS-CoV-2 has been reported. However, without RNA extraction the variplex test system failed to reliable detect SARS-CoV-2 directly in respiratory samples.

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Section: Discussionmentioning

confidence: 99%

“…The potential of LAMP-technology has been demonstrated impressively. Synthesized RNA of SARS-CoV-2 could be ampli ed to detectable levels in dilutions as low as 2 -100 copies per reaction [12,13].…”

Section: Discussionmentioning

confidence: 99%

VariplexTM test system fails to reliably detect SARS-CoV-2 directly from respiratory samples without RNA extraction

Eckel

Küsters

Drossel

et al. 2020

Preprint

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“…Revisions and iterations were made frequently to improve the functionality, usability and practicality. Technical details on the development of the rapid test kit, including clinical validation, can be found in our previous paper [2].…”

Section: Testing Iteration and Optimisationmentioning

confidence: 99%

“…We have developed a rapid test kit for detecting SARS-CoV-2 that can be performed on field without the need for a laboratory or specialist equipment. The technical details have been reported elsewhere [2]. Here, we present the methodol-ogy behind the design and development of this test kit as an example of a product design exercise.…”

Section: Introductionmentioning

confidence: 99%

Development of a rapid test kit for SARS-CoV-2: an example of product design

et al. 2020

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We present an example of applying 'need-driven' product design principle to the development of a rapid test kit to detect SARS-COV-2 (COVID-19). The tests are intended for use in the field and, longer term, for home use. They detect whether a subject is currently infected with the virus and is infectious. The urgent need for large numbers of tests in field setting imposes constraints such as short test time and lack of access to specialist equipment, laboratories and skilled technicians to perform the test and interpret results. To meet these needs, an antigen test based on RT-LAMP with colorimetric readout was chosen. Direct use of swab sample with no RNA extraction was explored. After extensive experimental study (reported elsewhere), a rapid test kit has been fabricated to satisfy all design criteria.

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“…Since the first reports of the SARS-CoV-2 virus 2 several studies have proposed new diagnostic approaches that make use of frontier technologies to detect SARS-CoV-2. These include ultra-high-throughput methods based on next-generation sequencing 3,4 , highly informative methods using long-read sequencing 5 , molecular assays that reduce the need for laboratory infrastructure such as loopbased isothermal amplification (LAMP) [6][7][8] , CRISPR-Cas13 assays [9][10][11] , and immunology tests that detect the presence of anti-SARS-CoV-2 antibodies 12,13 . These technologies may increase the efficiency and accessibility of SARS-CoV-2 testing in the future but will require regulatory approval.…”

Section: Introductionmentioning

confidence: 99%

CONTAIN: An open-source shipping container laboratory optimised for automated COVID-19 diagnostics

Walker¹,

Donora²,

Thomas³

et al. 2020

Preprint

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The COVID-19 pandemic has challenged diagnostic systems globally. Expanding testing capabilities to conduct population-wide screening for COVID-19 requires innovation in diagnostic services at both the molecular and industrial scale. No report to-date has considered the complexity of laboratory infrastructure in conjunction with the available molecular assays to offer a standardised solution to testing. Here we present CONTAIN. A modular biosafety level 2+ laboratory optimised for automated testing based on a standard 40ft shipping container. Using open-source liquid-handling robots and RNA extraction reagents we demonstrate a reproducible workflow for RT-qPCR COVID-19 testing. With five OT2 liquid handlers, a single CONTAIN unit reaches a maximum daily testing capacity of 2400 tests/day. We validate this workflow for automated RT-qPCR testing, using both synthetic SARS-CoV-2 samples and patient samples from a local NHS hospital. Finally, we discuss the suitability of CONTAIN and its flexibility in a range of diagnostic testing scenarios including high-density urban environments and mobile response units. Visual abstractCONTAIN: An open-source shipping container laboratory optimised for automated COVID-19 diagnostics

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RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2

Cited by 459 publications

References 26 publications

VariplexTM test system fails to reliably detect SARS-CoV-2 directly from respiratory samples without RNA extraction

VariplexTM test system fails to reliably detect SARS-CoV-2 directly from respiratory samples without RNA extraction

Development of a rapid test kit for SARS-CoV-2: an example of product design

CONTAIN: An open-source shipping container laboratory optimised for automated COVID-19 diagnostics

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