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Clinical Validation of a Rapid Variant-Proof RT-RPA Assay for the Detection of SARS-CoV-2.
Cherkaoui, Dounia; Heaney, Judith; Huang, Da; Byott, Matthew; Miller, Benjamin S; Nastouli, Eleni; McKendry, Rachel A.
  • Cherkaoui D; London Centre for Nanotechnology, University College London, London WC1E 6BT, UK.
  • Heaney J; Division of Medicine, University College London, London WC1E 6BT, UK.
  • Huang D; Advanced Pathogen Diagnostics Unit, University College London Hospitals (UCLH) NHS Trust, London NW1 2BU, UK.
  • Byott M; London Centre for Nanotechnology, University College London, London WC1E 6BT, UK.
  • Miller BS; Advanced Pathogen Diagnostics Unit, University College London Hospitals (UCLH) NHS Trust, London NW1 2BU, UK.
  • Nastouli E; Infection, Immunity and Inflammation Department, Great Ormond Street Institute for Child Health (ICH), University College London, London WC1N 1EH, UK.
  • McKendry RA; London Centre for Nanotechnology, University College London, London WC1E 6BT, UK.
Diagnostics (Basel) ; 12(5)2022 May 19.
Article in English | MEDLINE | ID: covidwho-1928513
ABSTRACT
The COVID-19 pandemic has unveiled a pressing need to expand the diagnostic landscape to permit high-volume testing in peak demand. Rapid nucleic acid testing based on isothermal amplification is a viable alternative to real-time reverse transcription polymerase chain reaction (RT-PCR) and can help close this gap. With the emergence of SARS-CoV-2 variants of concern, clinical validation of rapid molecular tests needs to demonstrate their ability to detect known variants, an essential requirement for a robust pan-SARS-CoV-2 assay. To date, there has been no clinical validation of reverse transcription recombinase polymerase amplification (RT-RPA) assays for SARS-CoV-2 variants. We performed a clinical validation of a one-pot multi-gene RT-RPA assay with the E and RdRP genes of SARS-CoV-2 as targets. The assay was validated with 91 nasopharyngeal samples, with a full range of viral loads, collected at University College London Hospitals. Moreover, the assay was tested with previously sequenced clinical samples, including eleven lineages of SARS-CoV-2. The rapid (20 min) RT-RPA assay showed high sensitivity and specificity, equal to 96% and 97%, respectively, compared to gold standard real-time RT-PCR. The assay did not show cross-reactivity with the panel of respiratory pathogens tested. We also report on a semi-quantitative analysis of the RT-RPA results with correlation to viral load equivalents. Furthermore, the assay could detect all eleven SARS-CoV-2 lineages tested, including four variants of concern (Alpha, Beta, Delta, and Omicron). This variant-proof SARS-CoV-2 assay offers a significantly faster and simpler alternative to RT-PCR, delivering sensitive and specific results with clinical samples.
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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Prognostic study / Randomized controlled trials Topics: Variants Language: English Year: 2022 Document Type: Article Affiliation country: Diagnostics12051263

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Prognostic study / Randomized controlled trials Topics: Variants Language: English Year: 2022 Document Type: Article Affiliation country: Diagnostics12051263