An Open One-Step RT-qPCR for SARS-CoV-2 detection

Simple item page
dc.catalogadoryvc
dc.contributor.authorCerda Rojas, Ariel Patricio
dc.contributor.authorRivera, Maira
dc.contributor.authorArmijo, Grace
dc.contributor.authorIbarra-Henríquez, Catalina
dc.contributor.authorReyes, Javiera
dc.contributor.authorBlázquez Sánchez, Paula
dc.contributor.authorAvilés, Javiera
dc.contributor.authorArce, Anibal
dc.contributor.authorSeguel, Aldo
dc.contributor.authorBrown, Alexander J.
dc.contributor.authorVásquez, Yesseny
dc.contributor.authorCortez-San Martín, Marcelo
dc.contributor.authorCubillos, Francisco A.
dc.contributor.authorGarcía, Patricia
dc.contributor.authorFerrés, Marcela
dc.contributor.authorRamírez Sarmiento, César Antonio
dc.contributor.authorFederici, Fernan
dc.contributor.authorGutiérrez, Rodrigo A.
dc.date.accessioned2025-03-12T16:50:25Z
dc.date.available2025-03-12T16:50:25Z
dc.date.issued2024
dc.description.abstractThe 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.
dc.description.funderANID Millennium Science Initiative Program (ICN17_022) to RAG, CARS and FF. Fondo de Desarrollo de Areas Prioritarias (Center for Genome Regulation; ANID FONDAP No. 15090007, to RAG and FF. Fondo de Desarrollo Científico y Tecnológico (FONDECYT 1201684 awarded to CARS, FONDECYT Regular No. 1211218 to FF, and FONDECYT No. 3190731 to MR; International Cooperation Program with Consejo Nacional de Ciencia, Tecnología e Innovación Tecnológica (ANID-CONCYTEC covbio 0012 awarded to FF and CARS). National Institutes of Health NIAID training grant (Training Program in Immunology; T32-AI07405) AJB. PBS and JR were supported by ANID Doctoral Scholarships (No. 21191979 and No. 21191684, respectively)
dc.fuente.origenWoS
dc.identifier.doi10.1371/journal.pone.0297081
dc.identifier.issn1932-6203
dc.identifier.urihttps://doi.org/10.1371/journal.pone.0297081
dc.identifier.urihttps://journals.plos.org/plosone/article?id=10.1371/journal.pone.0297081#abstract0
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/102530
dc.identifier.wosidWoS_ID: 001158471300068
dc.information.autorucFacultad de Ciencias Biológicas; Cerda Rojas, Ariel Patricio; S/I; 250244
dc.information.autorucInstituto de Ingeniería Biológica y Médica; Ramírez Sarmiento, César Antonio; 0000-0003-4647-903X; 1037905
dc.information.autorucFacultad de Ciencias Biológicas; Federici, Fernan; 0000-0001-9200-5383; 123357
dc.issue.numero1
dc.language.isoen
dc.nota.accesocontenido completo
dc.publisherPublic Library Science
dc.revistaPlos One
dc.rightsacceso abierto
dc.rights.licenseCC BY Atribución Internacional 4.0
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectSARS CoV 2
dc.subjectCOVID 19
dc.subjectVirus testing
dc.subjectReverse transcriptase-polymerase chain reaction
dc.subjectRibonucleases
dc.subjectEnzyme purification
dc.subjectRNA extraction
dc.subjectPolymerase chain reaction
dc.subject.ddc610
dc.subject.deweyMedicina y saludes_ES
dc.titleAn Open One-Step RT-qPCR for SARS-CoV-2 detection
dc.typeartículo
dc.volumen19
sipa.codpersvinculados250244
sipa.codpersvinculados1037905
sipa.codpersvinculados123357
sipa.indexWoS
sipa.trazabilidadWOS;2024-03-30
sipa.trazabilidadORCID;2025-03-03
Files
Collections
Artículos de revistas