ArticlePLoS biology2020
A sensitive and affordable multiplex RT-qPCR assay for SARS-CoV-2 detection.
Article in PLoS biology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed.
- Validation of Guanidine-EDTA as a Preservative Agent for the Analysis of miRNAs and mRNAs in Blood Samples of Chagas Disease Patients.Pathogens (Basel, Switzerland) · 2026Article
- Development and Validation of a Streamlined Multiplex RT-PCR Assay for Accurate SARS-CoV-2 Detection in Diverse Healthcare Environments.Current microbiology · 2026Article
- A Portable Dual-Mode Microfluidic Device Integrating RT-qPCR and RT-LAMP for Rapid Nucleic Acid Detection in Point-of-Care Testing.Biosensors · 2026Article
- Diagnostic value comparative analysis of the commercial kits for the detection of SARS-CoV-2 in clinical samples: a systematic review and meta-analysis.Iranian journal of microbiology · 2025Review
- High SARS-CoV-2 incidence and asymptomatic fraction during Delta and Omicron BA.1 waves in The Gambia.Nature communications · 2024Article
- An Open One-Step RT-qPCR for SARS-CoV-2 detection.PloS one · 2024Article
- Optimization and application of digital droplet PCR for the detection of SARS-CoV-2 in saliva specimen using commercially available kit.Biology methods & protocols · 2024Article
- Article
- The Many Mutations of the COVID-19 Variant: Current Perspectives on EG.5/Eris.Environmental health insights · 2023Review
- Mutations of SARS-CoV-2 and their impact on disease diagnosis and severity.Informatics in medicine unlocked · 2023Review
- Singleplex, multiplex and pooled sample real-time RT-PCR assays for detection of SARS-CoV-2 in an occupational medicine setting.Scientific reports · 2022Article
- User acceptability of saliva and gargle samples for identifying COVID-19 positive high-risk workers and household contacts.Diagnostic microbiology and infectious disease · 2022Article
- Factors associated with weak positive SARS-CoV-2 diagnosis by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR).Pathology · 2022Article
- Homebrew: Protocol for glassmilk-based nucleic-acid extraction for SARS-CoV-2 diagnostics.STAR protocols · 2022Article
- Duplex One-Step RT-qPCR Assays for Simultaneous Detection of Genomic and Subgenomic RNAs of SARS-CoV-2 Variants.Viruses · 2022Article
- Establishment of an in-house real-time RT-PCR assay for the detection of severe acute respiratory syndrome coronavirus 2 using the first World Health Organization international standard in a resource-limited country.Journal of clinical laboratory analysis · 2022Article
- Obtaining Reliable RT-qPCR Results in Molecular Diagnostics-MIQE Goals and Pitfalls for Transcriptional Biomarker Discovery.Life (Basel, Switzerland) · 2022Review
- New insights into the role of ribonuclease P protein subunit p30 from tumor to internal reference.Frontiers in oncology · 2022Review
- SARS-CoV-2 detection in wastewater using multiplex quantitative PCR.The Science of the total environment · 2021Article
- Implementation of an efficient SARS-CoV-2 specimen pooling strategy for high throughput diagnostic testing.Scientific reports · 2021Article
Corrections and comments
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Authors and funding
38 authors.
Funding
Abstract
With the ongoing COVID-19 (Coronavirus Disease 2019) pandemic, caused by the novel coronavirus SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2), there is a need for sensitive, specific, and affordable diagnostic tests to identify infected individuals, not all of whom are symptomatic. The most sensitive test involves the detection of viral RNA using RT-qPCR (quantitative reverse transcription PCR), with many commercial kits now available for this purpose. However, these are expensive, and supply of such kits in sufficient numbers cannot always be guaranteed. We therefore developed a multiplex assay using well-established SARS-CoV-2 targets alongside a human cellular control (RPP30) and a viral spike-in control (Phocine Herpes Virus 1 [PhHV-1]), which monitor sample quality and nucleic acid extraction efficiency, respectively. Here, we establish that this test performs as well as widely used commercial assays, but at substantially reduced cost. Furthermore, we demonstrate >1,000-fold variability in material routinely collected by combined nose and throat swabbing and establish a statistically significant correlation between the detected level of human and SARS-CoV-2 nucleic acids. The inclusion of the human control probe in our assay therefore provides a quantitative measure of sample quality that could help reduce false-negative rates. We demonstrate the feasibility of establishing a robust RT-qPCR assay at approximately 10% of the cost of equivalent commercial assays, which could benefit low-resource environments and make high-volume testing affordable.
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