ArticlePloS one2021
Extraction-free protocol combining proteinase K and heat inactivation for detection of SARS-CoV-2 by RT-qPCR.
Article in PloS one, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed.
- Development and Validation of a Streamlined Multiplex RT-PCR Assay for Accurate SARS-CoV-2 Detection in Diverse Healthcare Environments.Current microbiology · 2026Article
- The LAMP-CRISPR-Cas13a technique for detecting the CBASS mechanism of phage resistance in bacteria.Frontiers in microbiology · 2025Article
- Self-Diagnosis of SARS-CoV-2 from Saliva Samples at Home: Isothermal Amplification Enabled by Do-It-Yourself Portable Incubators and Laminated Poly-ethyl Sulfonate Membranes.Diagnostics (Basel, Switzerland) · 2024Article
- An Open One-Step RT-qPCR for SARS-CoV-2 detection.PloS one · 2024Article
- Extraction-Free Testing for SARS-CoV-2 in Nasal Swab and Saliva Samples on a Single High-Throughput Platform.Journal of biotechnology and biomedicine · 2024Article
- A novel viral RNA detection method based on the combined use of trans-acting ribozymes and HCR-FRET analyses.PloS one · 2024Article
- Optimization of extraction-free protocols for SARS-CoV-2 detection using a commercial rRT-PCR assay.Scientific reports · 2023Article
- Chemical Trends in Sample Preparation for Nucleic Acid Amplification Testing (NAAT): A Review.Biosensors · 2023Review
- Critical evaluation of strategies to achieve direct real-time PCR detection of swine pathogens in oral fluids.Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2023Article
- Evaluation of the AdvanSure One-Stop COVID-19 Plus Kit for SARS-CoV-2 Detection Using a Streamlined RNA Extraction Method.Annals of laboratory medicine · 2023Article
- CRISPR-Cas13a-Based Assay for Accurate Detection of OXA-48 and GES Carbapenemases.Microbiology spectrum · 2023Article
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- Replication in the presence of dengue convalescent serum impacts Zika virus neutralization sensitivity and fitness.Frontiers in cellular and infection microbiology · 2023Article
- Dry Swab-Based Nucleic Acid Extraction vs. Spin Column-Based Nucleic Acid Extraction for COVID-19 RT-PCR Testing: A Comparative Study.The Canadian journal of infectious diseases & medical microbiology = Journal canadien des maladies infectieuses et de la microbiologie medicale · 2023Article
- A Comprehensive Comparison of Rapid RNA Extraction Methods for Detection of SARS-CoV-2 as the Infectious Agent of the Upper Respiratory Tract using Direct RT-LAMP Assay.Advanced biomedical research · 2023Article
- Reverse Transcription-Loop-Mediated Isothermal Amplification-CRISPR-Cas13a Technology as a Promising Diagnostic Tool for SARS-CoV-2.Microbiology spectrum · 2022Article
- Pretreatment Methods for Human Nasopharyngeal Swabs to Increase the Signal to Noise Ratio of High Sensitivity Immunoassays.ACS measurement science au · 2022Article
- High-Throughput COVID-19 Testing of Naso-Oropharyngeal Swabs Using a Sensitive Extraction-Free Sample Preparation Method.Microbiology spectrum · 2022Article
- Strategies That Facilitate Extraction-Free SARS-CoV-2 Nucleic Acid Amplification Tests.Viruses · 2022Review
Corrections and comments
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Authors and funding
9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Real-time reverse transcription PCR (RT-qPCR) is the gold-standard technique for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection in nasopharyngeal swabs specimens. The analysis by RT-qPCR usually requires a previous extraction step to obtain the purified viral RNA. Unfortunately, RNA extraction constitutes a bottleneck for early detection in many countries since it is expensive, time-consuming and depends on the availability of commercial kits. Here, we describe an extraction-free protocol for SARS-CoV-2 detection by RT-qPCR from nasopharyngeal swab clinical samples in saline solution. The method includes a treatment with proteinase K followed by heat inactivation (PK+HID method). We demonstrate that PK+HID improves the RT-qPCR performance in comparison to the heat-inactivation procedure. Moreover, we show that this extraction-free protocol can be combined with a variety of multiplexing RT-qPCR kits. The method combined with a multiplexing detection kit targeting N and ORF1ab viral genes showed a sensitivity of 0.99 and a specificity of 0.99 from the analysis of 106 positive and 106 negative clinical samples. In conclusion, PK+HID is a robust, fast and inexpensive procedure for extraction-free RT-qPCR determinations of SARS-CoV-2. The National Administration of Drugs, Foods and Medical Devices of Argentina has recently authorized the use of this method.
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