ArticleHealthcare (Basel, Switzerland)2021
A Direct Method for RT-PCR Detection of SARS-CoV-2 in Clinical Samples.
Article in Healthcare (Basel, Switzerland), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Advances and Challenges in SARS-CoV-2 Detection: A Review of Molecular and Serological Technologies.Diagnostics (Basel, Switzerland) · 2024Review
- Deciphering the role of female reproductive tract microbiome in reproductive health: a review.Frontiers in cellular and infection microbiology · 2024Review
- Advances in the oral microbiota and rapid detection of oral infectious diseases.Frontiers in microbiology · 2023Review
- Article
- Rapid and Reliable Detection of SARS-CoV-2 Using Direct RT-LAMP.Diagnostics (Basel, Switzerland) · 2022Article
- Clinical profile, risk factors and outcomes of ric COVID-19: a retrospective cohort multicentre study in Saudi Arabia.BMJ open · 2022Observational
- Alternative RNA extraction-free techniques for the real-time RT-PCR detection of SARS-CoV-2 in nasopharyngeal swab and sputum samples.Journal of virological methods · 2021Article
- Science's Response to CoVID-19.ChemMedChem · 2021Review
- Enhanced rapid commercial DNA extraction kit for the molecular detection of severe acute respiratory syndrome coronavirus 2: Easy adaptation to current protocols.Revista da Sociedade Brasileira de Medicina Tropical · 2021Article
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
introductionthe emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global pandemic of acute respiratory disease (COVID-19). SARS-CoV-2 is a positive-strand RNA virus and its genomic characterization has played a vital role in the design of appropriate diagnostics tests. The current RT-PCR protocol for SARS-CoV-2 detects two regions of the viral genome, requiring RNA extraction and several hours. There is a need for fast, simple, and cost-effective detection strategies.
methodswe optimized a protocol for direct RT-PCR detection of SARS-CoV-2 without the need for nucleic acid extraction. Nasopharyngeal samples were diluted to 1:3 using diethyl pyrocarbonate (DEPC)-treated water. The diluted samples were incubated at 95 °C for 5 min in a thermal cycler, followed by a cooling step at 4 °C for 5 min. Samples then underwent reverse transcription real-time RT-PCR in the E and RdRp genes.
resultsour direct detection protocol showed 100% concordance with the standard protocol with an average Ct value difference of 4.38 for the E region and 3.85 for the RdRp region.
conclusionthe direct PCR technique was found to be a reliable and sensitive method that can be used to reduce the time and cost of the assay by removing the need for RNA extraction. It enables the use of the assay in research, diagnostics, and screening for COVID-19 in regions with fewer economic resources, where supplies are more limited allowing for wider use for screening.
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