Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection
Objectives: The ongoing COVID-19 pandemic continues to impose demands on diagnostic screening. In anticipation that the recurrence of outbreaks and the measures for lifting the lockdown worldwide may cause supply chain issues over the coming months, this study assessed the sensitivity of a number of...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-10-01
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| Series: | International Journal of Infectious Diseases |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S1201971220306032 |
| _version_ | 1818883023305703424 |
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| author | Julia Alcoba-Florez Helena Gil-Campesino Diego García-Martínez de Artola Rafaela González-Montelongo Agustín Valenzuela-Fernández Laura Ciuffreda Carlos Flores |
| author_facet | Julia Alcoba-Florez Helena Gil-Campesino Diego García-Martínez de Artola Rafaela González-Montelongo Agustín Valenzuela-Fernández Laura Ciuffreda Carlos Flores |
| author_sort | Julia Alcoba-Florez |
| collection | DOAJ |
| description | Objectives: The ongoing COVID-19 pandemic continues to impose demands on diagnostic screening. In anticipation that the recurrence of outbreaks and the measures for lifting the lockdown worldwide may cause supply chain issues over the coming months, this study assessed the sensitivity of a number of one-step retrotranscription and quantitative polymerase chain reaction (RT-qPCR) solutions to detect SARS-CoV-2. Methods: Six different RT-qPCR alternatives were evaluated for SARS-CoV-2/COVID-19 diagnosis based on standard RNA extractions. The one with best sensitivity was also assessed with direct nasopharyngeal swab viral transmission medium (VTM) heating; thus overcoming the RNA extraction step. Results: A wide variability in the sensitivity of RT-qPCR solutions was found that was associated with a range of false negatives from 2% (0.3–7.9%) to 39.8% (30.2–50.2%). Direct preheating of VTM combined with the best solution provided a sensitivity of 72.5% (62.5–81.0%), in the range of some of the solutions based on standard RNA extractions. Conclusions: Sensitivity limitations of currently used RT-qPCR solutions were found. These results will help to calibrate the impact of false negative diagnoses of COVID-19, and to detect and control new SARS-CoV-2 outbreaks and community transmissions. |
| first_indexed | 2024-12-19T15:27:03Z |
| format | Article |
| id | doaj.art-162dc79d88814442af166cf871150df9 |
| institution | Directory Open Access Journal |
| issn | 1201-9712 |
| language | English |
| last_indexed | 2024-12-19T15:27:03Z |
| publishDate | 2020-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | International Journal of Infectious Diseases |
| spelling | doaj.art-162dc79d88814442af166cf871150df92022-12-21T20:15:51ZengElsevierInternational Journal of Infectious Diseases1201-97122020-10-0199190192Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detectionJulia Alcoba-Florez0Helena Gil-Campesino1Diego García-Martínez de Artola2Rafaela González-Montelongo3Agustín Valenzuela-Fernández4Laura Ciuffreda5Carlos Flores6Servicio de Microbiología, Hospital Universitario N. S. de Candelaria, Santa Cruz de Tenerife, SpainServicio de Microbiología, Hospital Universitario N. S. de Candelaria, Santa Cruz de Tenerife, SpainServicio de Microbiología, Hospital Universitario N. S. de Candelaria, Santa Cruz de Tenerife, SpainGenomics Division, Instituto Tecnológico y de Energías Renovables, Santa Cruz de Tenerife, SpainLaboratorio de Inmunología Celular y Viral, Unidad de Farmacología, Facultad de Medicina & IUETSPC, Universidad de La Laguna, San Cristóbal de La Laguna, Spain; Red española de Investigación en VIH/SIDA (RIS)-RETIC, Instituto de Salud Carlos III, Madrid, SpainResearch Unit, Hospital Universitario N. S. de Candelaria, Santa Cruz de Tenerife, SpainGenomics Division, Instituto Tecnológico y de Energías Renovables, Santa Cruz de Tenerife, Spain; Research Unit, Hospital Universitario N. S. de Candelaria, Santa Cruz de Tenerife, Spain; CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, Madrid, Spain; Instituto de Tecnologías Biomédicas (ITB) Universidad de La Laguna, San Cristóbal de La Laguna, Spain; Corresponding author at: Unidad de Investigación, Hospital Universitario N.S. de Candelaria, Carretera del Rosario s/n, 38010 Santa Cruz de Tenerife, Spain.Objectives: The ongoing COVID-19 pandemic continues to impose demands on diagnostic screening. In anticipation that the recurrence of outbreaks and the measures for lifting the lockdown worldwide may cause supply chain issues over the coming months, this study assessed the sensitivity of a number of one-step retrotranscription and quantitative polymerase chain reaction (RT-qPCR) solutions to detect SARS-CoV-2. Methods: Six different RT-qPCR alternatives were evaluated for SARS-CoV-2/COVID-19 diagnosis based on standard RNA extractions. The one with best sensitivity was also assessed with direct nasopharyngeal swab viral transmission medium (VTM) heating; thus overcoming the RNA extraction step. Results: A wide variability in the sensitivity of RT-qPCR solutions was found that was associated with a range of false negatives from 2% (0.3–7.9%) to 39.8% (30.2–50.2%). Direct preheating of VTM combined with the best solution provided a sensitivity of 72.5% (62.5–81.0%), in the range of some of the solutions based on standard RNA extractions. Conclusions: Sensitivity limitations of currently used RT-qPCR solutions were found. These results will help to calibrate the impact of false negative diagnoses of COVID-19, and to detect and control new SARS-CoV-2 outbreaks and community transmissions.http://www.sciencedirect.com/science/article/pii/S1201971220306032COVID-19SARS-CoV-2DiagnosisFalse negativesSolution comparisonsSensitivity |
| spellingShingle | Julia Alcoba-Florez Helena Gil-Campesino Diego García-Martínez de Artola Rafaela González-Montelongo Agustín Valenzuela-Fernández Laura Ciuffreda Carlos Flores Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection International Journal of Infectious Diseases COVID-19 SARS-CoV-2 Diagnosis False negatives Solution comparisons Sensitivity |
| title | Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection |
| title_full | Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection |
| title_fullStr | Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection |
| title_full_unstemmed | Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection |
| title_short | Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection |
| title_sort | sensitivity of different rt qpcr solutions for sars cov 2 detection |
| topic | COVID-19 SARS-CoV-2 Diagnosis False negatives Solution comparisons Sensitivity |
| url | http://www.sciencedirect.com/science/article/pii/S1201971220306032 |
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