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Capillary-based reverse transcriptase loop-mediated isothermal amplification for cost-effective and rapid point-of-care COVID-19 testing

Capillary-based reverse transcriptase loop-mediated isothermal amplification for cost-effective and rapid point-of-care COVID-19 testing
Capillary-based reverse transcriptase loop-mediated isothermal amplification for cost-effective and rapid point-of-care COVID-19 testing
As the SARS-CoV-2 pandemic continues to spread, the necessity for rapid, easy diagnostic capabilities could never have been more crucial. With this aim in mind, we have developed a cost-effective and time-saving testing
methodology/strategy that implements a sensitive reverse transcriptase loop-mediated amplification (RT-LAMP) assay within narrow, commercially available and cheap, glass capillaries for detection of the SARS-CoV-2 viral
RNA. The methodology is compatible with widely used laboratory-based molecular testing protocols and currently available infrastructure. It employs a simple rapid extraction protocol that lyses the virus, releasing sufficient
genetic material for amplification. This extracted viral RNA is then amplified using a SARS-CoV-2 RT-LAMP
kit, at a constant temperature and the resulting amplified product produces a colour change which can be visually interpreted. This testing protocol, in conjunction with the RT-LAMP assay, has a sensitivity of ∼100 viral
copies per reaction of a sample and provides results in a little over 30 min. As the assay is carried out in a water bath, commonly available within most testing laboratories, it eliminates the need for specialised instruments and
associated skills. In addition, our testing pathway requires a significantly reduced quantity of reagents per test while providing comparable sensitivity and specificity to the RT-LAMP kit used in this study. While the conventional
technique requires 25 μl of reagent, our test only utilises less than half the quantity (10 μl). Thus, with its minimalistic approach, this capillary-based assay could be a promising alternative to the conventional testing, owing to the fact that it can be performed in resource-limited settings, using readily available apparatus, and has the potential of increasing the overall testing capacity, while also reducing the burden on supply chains for mass
testing.
COVID-19, Capillary polymerase chain reaction (PCR), Loop-mediated isothermal amplification (LAMP), Molecular diagnostic
0003-2670
John, Anto
518d3739-712a-4681-9d0c-3e4fccf00454
He, Peijun
6faacea8-db2e-4bab-8198-3aa595724794
Katis, Ioannis
f92dfb8f-610d-4877-83f6-fd26a571df12
Galanis, Panagiotis
4457b788-deef-4293-ab39-76f501b9529d
Iles, Alice
4c7e0b28-8a6e-442e-88ca-a06e6e1a453d
Eason, R.W.
e38684c3-d18c-41b9-a4aa-def67283b020
Sones, Collin
9de9d8ee-d394-46a5-80b7-e341c0eed0a8
John, Anto
518d3739-712a-4681-9d0c-3e4fccf00454
He, Peijun
6faacea8-db2e-4bab-8198-3aa595724794
Katis, Ioannis
f92dfb8f-610d-4877-83f6-fd26a571df12
Galanis, Panagiotis
4457b788-deef-4293-ab39-76f501b9529d
Iles, Alice
4c7e0b28-8a6e-442e-88ca-a06e6e1a453d
Eason, R.W.
e38684c3-d18c-41b9-a4aa-def67283b020
Sones, Collin
9de9d8ee-d394-46a5-80b7-e341c0eed0a8

John, Anto, He, Peijun, Katis, Ioannis, Galanis, Panagiotis, Iles, Alice, Eason, R.W. and Sones, Collin (2021) Capillary-based reverse transcriptase loop-mediated isothermal amplification for cost-effective and rapid point-of-care COVID-19 testing. Analytica Chimica Acta, 1185, [339002]. (doi:10.1016/j.aca.2021.339002).

Record type: Article

Abstract

As the SARS-CoV-2 pandemic continues to spread, the necessity for rapid, easy diagnostic capabilities could never have been more crucial. With this aim in mind, we have developed a cost-effective and time-saving testing
methodology/strategy that implements a sensitive reverse transcriptase loop-mediated amplification (RT-LAMP) assay within narrow, commercially available and cheap, glass capillaries for detection of the SARS-CoV-2 viral
RNA. The methodology is compatible with widely used laboratory-based molecular testing protocols and currently available infrastructure. It employs a simple rapid extraction protocol that lyses the virus, releasing sufficient
genetic material for amplification. This extracted viral RNA is then amplified using a SARS-CoV-2 RT-LAMP
kit, at a constant temperature and the resulting amplified product produces a colour change which can be visually interpreted. This testing protocol, in conjunction with the RT-LAMP assay, has a sensitivity of ∼100 viral
copies per reaction of a sample and provides results in a little over 30 min. As the assay is carried out in a water bath, commonly available within most testing laboratories, it eliminates the need for specialised instruments and
associated skills. In addition, our testing pathway requires a significantly reduced quantity of reagents per test while providing comparable sensitivity and specificity to the RT-LAMP kit used in this study. While the conventional
technique requires 25 μl of reagent, our test only utilises less than half the quantity (10 μl). Thus, with its minimalistic approach, this capillary-based assay could be a promising alternative to the conventional testing, owing to the fact that it can be performed in resource-limited settings, using readily available apparatus, and has the potential of increasing the overall testing capacity, while also reducing the burden on supply chains for mass
testing.

Text
pagination_ACA_339002 (5) - Accepted Manuscript
Restricted to Repository staff only until 31 August 2022.
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More information

Accepted/In Press date: 26 August 2021
e-pub ahead of print date: 31 August 2021
Keywords: COVID-19, Capillary polymerase chain reaction (PCR), Loop-mediated isothermal amplification (LAMP), Molecular diagnostic

Identifiers

Local EPrints ID: 452274
URI: http://eprints.soton.ac.uk/id/eprint/452274
ISSN: 0003-2670
PURE UUID: 5d443e12-2175-4eb1-b40c-ceef5e466224
ORCID for Ioannis Katis: ORCID iD orcid.org/0000-0002-2016-557X
ORCID for Panagiotis Galanis: ORCID iD orcid.org/0000-0002-2028-5803
ORCID for R.W. Eason: ORCID iD orcid.org/0000-0001-9704-2204

Catalogue record

Date deposited: 02 Dec 2021 17:34
Last modified: 28 Apr 2022 02:21

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Contributors

Author: Anto John
Author: Peijun He
Author: Ioannis Katis ORCID iD
Author: Panagiotis Galanis ORCID iD
Author: Alice Iles
Author: R.W. Eason ORCID iD
Author: Collin Sones

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