| Author | Queiroz, Jackson Alves da Silva | |
| Author | Rampazzo, Rita de Cássia Pontello | |
| Author | Silva Filho, Edivá Basílio da | |
| Author | Oliveira, Gabriella Sgorlon | |
| Author | Oliveira, Suyane da Costa | |
| Author | Souza, Luan Felipo Botelho | |
| Author | Pereira, Soraya dos Santos | |
| Author | Rodrigues, Moreno Magalhães de Souza | |
| Author | Maia, Adriana Cristina Salvador | |
| Author | Silva, Cicileia Correia da | |
| Author | Mendonça, Aline Linhares Ferreira de Melo | |
| Author | Lugtenburg, Celina Aparecida Bertoni | |
| Author | Aguiar, Francisco de Assis Araújo | |
| Author | Rodrigues, Rosiane de Souza Soares | |
| Author | Santos, Caio Henrique Nemeth | |
| Author | Guimarães, Alice Paula Di Sabatino | |
| Author | Máximo, Fernando Rodrigues | |
| Author | Santos, Alcione de Oliveira dos | |
| Author | Krieger, Marco Aurélio | |
| Author | Salcedo, Juan Miguel Villalobos | |
| Author | Dall’Acqua, Deusilene Souza Vieira | |
| Access date | 2021-02-03T13:49:56Z | |
| Available date | 2021-02-03T13:49:56Z | |
| Document date | 2021 | |
| Citation | QUEIROZ, Jackson Alves da Silva et al. Development of a quantitative one-step multiplex RT-qPCR assay for the detection of SARS-CoV-2 in a biological matrix. International Journal of Infectious Diseases, p. 1-21, 1 Jan. 2021. | |
| ISSN | 1201-9712 | |
| URI | https://www.arca.fiocruz.br/handle/icict/45943 | |
| Language | eng | en_US |
| Publisher | Elsevier | |
| Later version | https://www.arca.fiocruz.br/handle/icict/46315 | |
| Rights | open access | |
| Title | Development of a quantitative one-step multiplex RT-qPCR assay for the detection of SARS-CoV-2 in a biological matrix (preprint) | en_US |
| Type | Preprint | |
| DOI | 10.1016/j.ijid.2021.01.001 | |
| Abstract | Introduction: COVID-19 is a disease caused by Severe Acute Respiratory Syndrome Virus 2 (SARS-CoV-2) which emerged in China in late 2019. The rapid viral spread has made the disease a public health emergency of worldwide concern. The gold standard for diagnosing SARS-CoV-2 is reverse transcription followed by qualitative real-time polymerase chain reaction (RT-qPCR); however, the role of viral load quantification has not been thoroughly investigated yet. Objective: The aim of this study was to develop a high-precision quantitative one-step RT-qPCR reaction using the association of the viral target and the human target in the same reaction. Methods: The assay standardization involved the absolute quantification method with serial dilutions of a plasmid with the N gene in a biological matrix to build a standard curve. Results and Discussion: The results demonstrated the possibility of quantifying as few as 2.5 copies/reaction and analysis of 244 patients with known results selected by cross-section revealed 100% agreement with a qualitative RT-qPCR assay registered by Anvisa. In this population, it was possible to quantify patients with between 2.59 and 3.5x10⁷ copies per reaction and negative patients continued to indicate the same result. Conclusion: This assay can be a useful tool for the proper patients management, since the level and duration of viral replication are important factors to assess the risk of transmission and to guide decisions regarding the isolation and release of patients also, an accurate diagnosis is critical information whereas the current COVID-19 pandemic represents the biggest current global health problem. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil / Fundação Universidade Federal de Rondônia. Programa de Pós Graduação em Biologia Experimental. Porto Velho, RO, Brazil. | |
| Affilliation | Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brazil. | |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil / Fundação Universidade Federal de Rondônia. Programa de Pós Graduação em Biologia Experimental. Porto Velho, RO, Brazil. | |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Laboratório Central de Saúde Pública do Estado de Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil / Fundação Universidade Federal de Rondônia. Programa de Pós Graduação em Biologia Experimental. Porto Velho, RO, Brazil. | |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Laboratório Central de Saúde Pública do Estado de Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Laboratório Central de Saúde Pública do Estado de Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Laboratório Central de Saúde Pública do Estado de Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Laboratório Central de Saúde Pública do Estado de Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Laboratório Central de Saúde Pública do Estado de Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Laboratório Central de Saúde Pública do Estado de Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Secretaria de Estado de Finanças de Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Secretaria de Estado da Saúde de Rondônia. Porto Velho, RO, Brasil. | |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brazil. | |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil. | |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brazil / Fundação Universidade Federal de Rondônia. Programa de Pós Graduação em Biologia Experimental. Porto Velho, RO, Brazil. | |
| Subject | SARS-CoV-2 | en_US |
| Subject | Coronavirus Infections | en_US |
| Subject | Real-Time Polymerase Chain Reaction | en_US |
| Subject | Reagent Kits Diagnostic | en_US |
| Subject | COVID-19 | en_US |
