LoCKAmp: Lab-on-PCB technology for <3 minute virus genetic detection

Sotirios Papamatthaiou; James Boxall-Clasby; Ed Douglas; Pawel  Jajesniak; Hadrien  Peyret; June Mercer-Chalmers; Varun  K. S. Kumar; George  P. Lomonossoff; Julien Reboud; Maisem Laabei; Jonathan M. Cooper; Barbara Kasprzyk-Hordern; Despina Moschou

doi:10.1039/D3LC00441D

LoCKAmp: Lab-on-PCB technology for <3 minute virus genetic detection

Sotirios Papamatthaiou, James Boxall-Clasby, Ed Douglas, Pawel Jajesniak, Hadrien Peyret, June Mercer-Chalmers, Varun K. S. Kumar, George P. Lomonossoff, Julien Reboud, Maisem Laabei, Jonathan M. Cooper, Barbara Kasprzyk-Hordern, Despina Moschou

Research output: Contribution to journal › Article › peer-review

6 Citations (SciVal)

Abstract

The recent COVID-19 outbreak highlighted the need for lab-on-chip diagnostic technology fit for real-life deployment in the field. Existing bottlenecks in multistep analytical microsystem integration and upscalable, standardized fabrication techniques delayed the large-scale deployment of lab-on-chip solutions during the outbreak, throughout a global diagnostic test shortage. This study presents a technology that has the potential to address these issues by redeploying and repurposing the ubiquitous printed circuit board (PCB) technology and manufacturing infrastructure. We demonstrate the first commercially manufactured, miniaturised lab-on-PCB device for loop-mediated isothermal amplification (LAMP) genetic detection of SARS-CoV-2. The system incorporates a mass-manufactured, continuous-flow PCB chip with ultra-low cost fluorescent detection circuitry, rendering it the only continuous-flow μLAMP platform with off-the-shelf optical detection components. Ultrafast, SARS-CoV-2 RNA amplification in wastewater samples was demonstrated within 2 min analysis, at concentrations as low as 17 gc μL ⁻¹. We further demonstrate our device operation by detecting SARS-CoV-2 in 20 human nasopharyngeal swab samples, without the need for any RNA extraction or purification. This renders the presented miniaturised nucleic-acid amplification-based diagnostic test the fastest reported SARS-CoV-2 genetic detection platform, in a practical implementation suitable for deployment in the field. This technology can be readily extended to the detection of alternative pathogens or genetic targets for a very broad range of applications and matrices. LoCKAmp lab-on-PCB chips are currently mass-manufactured in a commercial, ISO-compliant PCB factory, at a small-scale production cost of £2.50 per chip. Thus, with this work, we demonstrate a high technology-readiness-level lab-on-chip-based genetic detection system, successfully benchmarked against standard analytical techniques both for wastewater and nasopharyngeal swab SARS-CoV-2 detection.

Original language	English
Pages (from-to)	4400-4412
Number of pages	13
Journal	Lab on a Chip
Volume	23
Issue number	20
Early online date	7 Sept 2023
DOIs	https://doi.org/10.1039/D3LC00441D
Publication status	Published - 21 Oct 2023

Bibliographical note

Funding Information:
The authors at Department of Electronic and Electrical Engineering, University of Bath acknowledge financial support from Global Challenges Research Fund (GCRF) QR – UKRI and EPSRC IAA. At the John Innes Centre, this work was supported by the United Kingdom Biotechnology and Biological Sciences Research Council (BBSRC) Grant BB/V009087/1, the Institute Strategic Programme Grant “Molecules from Nature—Enhanced Research Capacity” (BBS/E/J/000PR9794), and the John Innes Foundation. The authors at Department of Biology & Biochemistry, University of Bath acknowledge financial support from the Academy of Medical Sciences (SBF006\1023). The authors would like to thank Mike Linham for his help in prototyping the microfluidic interface and the Royal United Hospitals Bath NHS Foundation Trust for the clinical sample collection.

Funding Information:
The authors at Department of Electronic and Electrical Engineering, University of Bath acknowledge financial support from Global Challenges Research Fund (GCRF) QR - UKRI and EPSRC IAA. At the John Innes Centre, this work was supported by the United Kingdom Biotechnology and Biological Sciences Research Council (BBSRC) Grant BB/V009087/1, the Institute Strategic Programme Grant “Molecules from Nature—Enhanced Research Capacity” (BBS/E/J/000PR9794), and the John Innes Foundation. The authors at Department of Biology & Biochemistry, University of Bath acknowledge financial support from the Academy of Medical Sciences (SBF006\1023). The authors would like to thank Mike Linham for his help in prototyping the microfluidic interface and the Royal United Hospitals Bath NHS Foundation Trust for the clinical sample collection.

Publisher Copyright:
© 2023 The Royal Society of Chemistry.

Access to Document

10.1039/D3LC00441DLicence: CC BY

Cite this

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title = "LoCKAmp: Lab-on-PCB technology for <3 minute virus genetic detection",

abstract = "The recent COVID-19 outbreak highlighted the need for lab-on-chip diagnostic technology fit for real-life deployment in the field. Existing bottlenecks in multistep analytical microsystem integration and upscalable, standardized fabrication techniques delayed the large-scale deployment of lab-on-chip solutions during the outbreak, throughout a global diagnostic test shortage. This study presents a technology that has the potential to address these issues by redeploying and repurposing the ubiquitous printed circuit board (PCB) technology and manufacturing infrastructure. We demonstrate the first commercially manufactured, miniaturised lab-on-PCB device for loop-mediated isothermal amplification (LAMP) genetic detection of SARS-CoV-2. The system incorporates a mass-manufactured, continuous-flow PCB chip with ultra-low cost fluorescent detection circuitry, rendering it the only continuous-flow μLAMP platform with off-the-shelf optical detection components. Ultrafast, SARS-CoV-2 RNA amplification in wastewater samples was demonstrated within 2 min analysis, at concentrations as low as 17 gc μL −1. We further demonstrate our device operation by detecting SARS-CoV-2 in 20 human nasopharyngeal swab samples, without the need for any RNA extraction or purification. This renders the presented miniaturised nucleic-acid amplification-based diagnostic test the fastest reported SARS-CoV-2 genetic detection platform, in a practical implementation suitable for deployment in the field. This technology can be readily extended to the detection of alternative pathogens or genetic targets for a very broad range of applications and matrices. LoCKAmp lab-on-PCB chips are currently mass-manufactured in a commercial, ISO-compliant PCB factory, at a small-scale production cost of £2.50 per chip. Thus, with this work, we demonstrate a high technology-readiness-level lab-on-chip-based genetic detection system, successfully benchmarked against standard analytical techniques both for wastewater and nasopharyngeal swab SARS-CoV-2 detection.",

author = "Sotirios Papamatthaiou and James Boxall-Clasby and Ed Douglas and Pawel Jajesniak and Hadrien Peyret and June Mercer-Chalmers and {K. S. Kumar}, Varun and {P. Lomonossoff}, George and Julien Reboud and Maisem Laabei and Cooper, {Jonathan M.} and Barbara Kasprzyk-Hordern and Despina Moschou",

note = "Funding Information: The authors at Department of Electronic and Electrical Engineering, University of Bath acknowledge financial support from Global Challenges Research Fund (GCRF) QR – UKRI and EPSRC IAA. At the John Innes Centre, this work was supported by the United Kingdom Biotechnology and Biological Sciences Research Council (BBSRC) Grant BB/V009087/1, the Institute Strategic Programme Grant “Molecules from Nature—Enhanced Research Capacity” (BBS/E/J/000PR9794), and the John Innes Foundation. The authors at Department of Biology & Biochemistry, University of Bath acknowledge financial support from the Academy of Medical Sciences (SBF006\1023). The authors would like to thank Mike Linham for his help in prototyping the microfluidic interface and the Royal United Hospitals Bath NHS Foundation Trust for the clinical sample collection. Funding Information: The authors at Department of Electronic and Electrical Engineering, University of Bath acknowledge financial support from Global Challenges Research Fund (GCRF) QR - UKRI and EPSRC IAA. At the John Innes Centre, this work was supported by the United Kingdom Biotechnology and Biological Sciences Research Council (BBSRC) Grant BB/V009087/1, the Institute Strategic Programme Grant “Molecules from Nature—Enhanced Research Capacity” (BBS/E/J/000PR9794), and the John Innes Foundation. The authors at Department of Biology & Biochemistry, University of Bath acknowledge financial support from the Academy of Medical Sciences (SBF006\1023). The authors would like to thank Mike Linham for his help in prototyping the microfluidic interface and the Royal United Hospitals Bath NHS Foundation Trust for the clinical sample collection. Publisher Copyright: {\textcopyright} 2023 The Royal Society of Chemistry.",

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AU - Jajesniak, Pawel

AU - Peyret, Hadrien

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LoCKAmp: Lab-on-PCB technology for <3 minute virus genetic detection

Abstract

Bibliographical note

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Dissecting complement resistance in Staphylococcus aureus

UKRI GCRF/Newton Fund : Building an Early Warning System for community-wide infectious disease spread: SARS-CoV-2 tracking in Africa via environment fingerprinting- COVID-19

Cite this

LoCKAmp: Lab-on-PCB technology for <3 minute virus genetic detection

Abstract

Bibliographical note

Access to Document

Other files and links

Fingerprint

Projects

Dissecting complement resistance in Staphylococcus aureus

UKRI GCRF/Newton Fund : Building an Early Warning System for community-wide infectious disease spread: SARS-CoV-2 tracking in Africa via environment fingerprinting- COVID-19

Cite this