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. 2020 Oct:99:190-192.
doi: 10.1016/j.ijid.2020.07.058. Epub 2020 Aug 1.

Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection

Julia Alcoba-Florez  1 , Helena Gil-Campesino  1 , Diego García-Martínez de Artola  1 , Rafaela González-Montelongo  2 , Agustín Valenzuela-Fernández  3 , Laura Ciuffreda  4 , Carlos Flores  5
Affiliations

Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection

Julia Alcoba-Florez et al. Int J Infect Dis. 2020 Oct.

Abstract

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.

Keywords: COVID-19; Diagnosis; False negatives; SARS-CoV-2; Sensitivity; Solution comparisons.

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Figures

Fig. 1
Fig. 1
Raincloud plot of the distribution of cycle threshold (Ct) values for the RT-qPCR solutions evaluated for the detection of SARS-CoV-2 in COVID-19 positive samples. Raw Ct data with the median and the interquartile range are also represented and overlaid on each distribution.
See this image and copyright information in PMC

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