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

Autor:	Cerda A; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; FONDAP Center for Genome Regulation. Departamento de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago, 8331150, Chile., Rivera M; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; Institute for Biological and Medical Engineering, Schools of Engineering, Medicine and Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile., Armijo G; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; FONDAP Center for Genome Regulation. Departamento de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago, 8331150, Chile., Ibarra-Henriquez C; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; FONDAP Center for Genome Regulation. Departamento de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago, 8331150, Chile., Reyes J; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; Institute for Biological and Medical Engineering, Schools of Engineering, Medicine and Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile., Blázquez-Sánchez P; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; Institute for Biological and Medical Engineering, Schools of Engineering, Medicine and Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile., Avilés J; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio)., Arce A; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio)., Seguel A; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio)., Brown AJ; Department of Biomedical Research, National Jewish Health, Denver, CO, USA.; Department of Immunology & Microbiology, University of Colorado Anschutz Medical Campus, Aurora, CO, USA., Vásquez Y; Escuela de Ciencias Médicas. Facultad de Medicina. Universidad de Santiago de Chile. USACH, Santiago, Chile., Cortez-San Martín M; Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, USACH, Santiago, Chile., Cubillos FA; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, USACH, Santiago, Chile., García P; Departamento de Laboratorios Clínicos. Escuela de Medicina. Facultad de Medicina. Pontificia Universidad Católica de Chile, Santiago, Chile., Ferres M; Departamento de Laboratorios Clínicos. Escuela de Medicina. Facultad de Medicina. Pontificia Universidad Católica de Chile, Santiago, Chile., Ramírez-Sarmiento CA; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; Institute for Biological and Medical Engineering, Schools of Engineering, Medicine and Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile., Federici F; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; FONDAP Center for Genome Regulation. Departamento de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago, 8331150, Chile.; Institute for Biological and Medical Engineering, Schools of Engineering, Medicine and Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile., Gutiérrez RA; ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).; FONDAP Center for Genome Regulation. Departamento de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago, 8331150, Chile.
Jazyk:	angličtina
Zdroj:	MedRxiv : the preprint server for health sciences [medRxiv] 2023 May 26. Date of Electronic Publication: 2023 May 26.
DOI:	10.1101/2021.11.29.21267000
Abstrakt:	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, and the kit showed comparable sensitivity to approved commercial kits. The One-Step RT-qPCR was then tested on clinical samples and demonstrated similar performance to commercial kits in terms of positive and negative calls. 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. Competing Interests: Competing interests The authors have declared that no competing interests exist.
Databáze:	MEDLINE
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