ArticleScientific reports2020
CoV2-ID, a MIQE-compliant sub-20-min 5-plex RT-PCR assay targeting SARS-CoV-2 for the diagnosis of COVID-19.
Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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19 citing papers in PubMed, 31 citations in OpenAlex.
- A proof of concept study on the diagnostic utility of in vivo expressed mycobacterial transcripts in tuberculous pleuritis.Scientific reports · 2026Article
- ƩS COVID-19 is a rapid high throughput and sensitive one-step quadruplex real-time RT-PCR assay.Scientific reports · 2024Article
- Nucleic Acid Based Testing (NABing): A Game Changer Technology for Public Health.Molecular biotechnology · 2024Review
- FlashPCR: Revolutionising qPCR by Accelerating Amplification through Low ∆T Protocols.International journal of molecular sciences · 2024Article
- SARS2Mutant: SARS-CoV-2 amino-acid mutation atlas database.NAR genomics and bioinformatics · 2023Article
- A Systematic Review and Meta-Analysis Comparing the Diagnostic Accuracy Tests of COVID-19.Diagnostics (Basel, Switzerland) · 2023Review
- Digital PCR Applications in the SARS-CoV-2/COVID-19 Era: a Roadmap for Future Outbreaks.Clinical microbiology reviews · 2022Review
- Digital PCR can augment the interpretation of RT-qPCR Cq values for SARS-CoV-2 diagnostics.Methods (San Diego, Calif.) · 2022Article
- RT-qPCR Detection of SARS-CoV-2: No Need for a Dedicated Reverse Transcription Step.International journal of molecular sciences · 2022Article
- Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance.The Science of the total environment · 2022Review
- Detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its first variants in fourplex real-time quantitative reverse transcription-PCR assays.Microbial cell (Graz, Austria) · 2022Article
- Comparison between the analytical sensitivity and clinical performance of two cobas SARS-CoV-2 tests based on high-throughput and point-of-care systems.BioMedicine · 2022Article
- Design and storage stability of reference materials for microfluidic quantitative PCR-based equine gene doping tests.Journal of equine science · 2021Article
- Article
- RT-qPCR Diagnostics: The "Drosten" SARS-CoV-2 Assay Paradigm.International journal of molecular sciences · 2021Article
- Sample Adequacy Control (SAC) Lowers False Negatives and Increases the Quality of Screening: Introduction of "Non-Competitive" SAC for qPCR Assays.Diagnostics (Basel, Switzerland) · 2021Article
- Development of a PDRA Method for Detection of the D614G Mutation in COVID-19 Virus - Worldwide, 2021.China CDC weekly · 2021Article
- COVID-19 and Diagnostic Testing for SARS-CoV-2 by RT-qPCR-Facts and Fallacies.International journal of molecular sciences · 2021Article
- Detecting SARS-CoV-2 Orf3a and E ion channel activity in COVID-19 blood samples.Journal of clinical and translational science · 2021Article
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Accurate, reliable and rapid detection of SARS-CoV-2 is essential not only for correct diagnosis of individual COVID-19 disease but also for the development of a rational strategy aimed at lifting confinement restrictions and preparing for possible recurrent waves of viral infections. We have used the MIQE guidelines to develop two versions of a unique five plex RT-qPCR test, termed CoV2-ID, that allows the detection of three viral target genes, a human internal control for confirming the presence of human cells in a sample and a control artificial RNA for quality assessment and potential quantification. Viral targets can be detected either individually with separate fluorophores or jointly using the same fluorophore, thus increasing the test's reliability and sensitivity. It is robust, can consistently detect two copies of viral RNA, with a limit of detection of a single copy and can be completed in around 15 min. It was 100% sensitive and 100% specific when tested on 23 RNA samples extracted from COVID-19 positive patients and five COVID-19 negative patients. We also propose using multiple cycle fluorescence detection, rather than real-time PCR to reduce significantly the time taken to complete the assay as well as assuage the misunderstandings underlying the use of quantification cycles (Cq). Finally, we have designed an assay for the detection of the D614G mutation and show that all of the samples isolated in the Chelmsford, Essex area between mid-April and June 2020, have the mutant genotype whereas a sample originating in Australia was infected with the wild type genotype.
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