An Open One-Step RT-qPCR for SARS-CoV-2 detection
The COVID-19 pandemic has resulted in millions of deaths globally, and while several diagnostic systems were proposed, real-time reverse transcription polymerase chain reaction (RT-PCR) remains the gold standard. However, diagnostic reagents, including enzymes used in RT-PCR, are subject to centrali...
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| Veröffentlicht in: | PloS one 2024-01, Vol.19 (1), p.e0297081 |
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| creator | Cerda, Ariel Rivera, Maira Armijo, Grace Ibarra-Henriquez, Catalina Reyes, Javiera Blázquez-Sánchez, Paula Avilés, Javiera Arce, Aníbal Seguel, Aldo Brown, Alexander J Vásquez, Yesseny Cortez-San Martín, Marcelo Cubillos, Francisco A García, Patricia Ferres, Marcela Ramírez-Sarmiento, César A Federici, Fernán Gutiérrez, Rodrigo A |
| description | The COVID-19 pandemic has resulted in millions of deaths globally, and while several diagnostic systems were proposed, real-time reverse transcription polymerase chain reaction (RT-PCR) remains the gold standard. However, diagnostic reagents, including enzymes used in RT-PCR, are subject to centralized production models and intellectual property restrictions, which present a challenge for less developed countries. With the aim of generating a standardized One-Step open RT-qPCR protocol to detect SARS-CoV-2 RNA in clinical samples, we purified and tested recombinant enzymes and a non-proprietary buffer. The protocol utilized M-MLV RT and Taq DNA pol enzymes to perform a Taqman probe-based assay. Synthetic RNA samples were used to validate the One-Step RT-qPCR components, demonstrating sensitivity comparable to a commercial kit routinely employed in clinical settings for patient diagnosis. Further evaluation on 40 clinical samples (20 positive and 20 negative) confirmed its comparable diagnostic accuracy. This study represents a proof of concept for an open approach to developing diagnostic kits for viral infections and diseases, which could provide a cost-effective and accessible solution for less developed countries. |
| doi_str_mv | 10.1371/journal.pone.0297081 |
| format | Article |
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This study represents a proof of concept for an open approach to developing diagnostic kits for viral infections and diseases, which could provide a cost-effective and accessible solution for less developed countries.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>38271448</pmid><doi>10.1371/journal.pone.0297081</doi><tpages>e0297081</tpages><orcidid>https://orcid.org/0000-0003-3232-909X</orcidid><orcidid>https://orcid.org/0000-0002-5961-5005</orcidid><orcidid>https://orcid.org/0000-0003-4647-903X</orcidid><orcidid>https://orcid.org/0000-0002-9638-7400</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2024-01, Vol.19 (1), p.e0297081 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_3069270444 |
| source | PubMed Central (Open Access); MEDLINE; Public Library of Science; Free Full-Text Journals in Chemistry; EZB Electronic Journals Library; Directory of Open Access Journals (Open Access) |
| subjects | Analysis Biology and life sciences Clinical Laboratory Techniques - methods Collaboration Confidentiality COVID-19 COVID-19 - diagnosis COVID-19 Testing Developed countries Diagnostic systems Disease transmission DNA DNA polymerase Enzymes Evaluation Humans Intellectual property Laboratories Medicine and health sciences Pandemics Pathogens Personal protective equipment Polymerase chain reaction Reagents Real time Research and Analysis Methods Reverse transcription Ribonucleic acid RNA RNA probes RNA, Viral - analysis RNA, Viral - genetics SARS-CoV-2 - genetics Sensitivity and Specificity Severe acute respiratory syndrome coronavirus 2 Supplies Viral diseases |
| title | An Open One-Step RT-qPCR for SARS-CoV-2 detection |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-14T15%3A56%3A00IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=An%20Open%20One-Step%20RT-qPCR%20for%20SARS-CoV-2%20detection&rft.jtitle=PloS%20one&rft.au=Cerda,%20Ariel&rft.date=2024-01-25&rft.volume=19&rft.issue=1&rft.spage=e0297081&rft.pages=e0297081-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0297081&rft_dat=%3Cgale_plos_%3EA780401160%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=3069270444&rft_id=info:pmid/38271448&rft_galeid=A780401160&rft_doaj_id=oai_doaj_org_article_6b559cda8fab4445abc7831909630f3d&rfr_iscdi=true |