Rapid establishment of laboratory diagnostics for the novel coronavirus SARS-CoV-2 in Bavaria, Germany, February 2020

Abstract

The need for timely establishment of diagnostic assays arose when Germany was confronted with the first travel-associated outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Europe. We describe our laboratory experiences during a large contact tracing investigation, comparing previously published real-time RT-PCR assays in different PCR systems and a commercial kit. We found that assay performance using the same primers and probes with different PCR systems varied and the commercial kit performed well.At the end of December 2019, an outbreak caused by a novel coronavirus was announced in Wuhan, China. Since then, the number of cases has increased, especially in China but also in other countries, and public health authorities are in need to rapidly implement diagnostic tools. In this paper, we describe our laboratory experiences with the novel real-time RT-PCR assays comparing different one-step PCR systems and a commercial kit, using a Bio-Rad CFX 96 cycler.

The epidemiology, diagnosis and treatment of COVID-19

“…Among the three novel assays, the COVID-19-RdRp/Hel assay had the lowest limit of detection in vitro; highly sensitive and specific assays may help to improve the laboratory diagnosis of COVID-19 [20] . The SARS-CoV E gene assay was more sensitive than the RdRp gene assay combined with the one-step RT-PCR system [21] . The E gene PCR was sufficient to diagnose a SARS-CoV-2 infection but the RdRp protocol was recommended to confirm a positive result [22 , 23] .…”

COVID-19: A promising cure for the global panic

“…Contrastingly, the E protein with RdRp was also detecting SARS-CoV, and so, these assays can be used to test for the SARS-CoV-2 when there are no traces of SARS-CoV (Cordes and Heim, 2020). When the commercially available Real Star kit, Virus +Rox Vial kit and Super Script III Onestep RT-PCR System with Platinum TaqDNA Polymerase were compared for their efficiency, the RealStar Kit did not have any unwanted signals and exceeded the other two in its performance (Konrad et al, 2020). These methods can also be compromised due to inadequate sample volume, inaccuracies in methods of testing, not collecting samples at the appropriate time window, and contamination.…”

An alternative workflow for molecular detection of SARS-CoV-2 - escape from the NA extraction kit-shortage, Copenhagen, Denmark, March 2020

“…at 4˚C before being used in the RT-qPCR reaction. Two SARS-CoV-2 RT-qPCR assays were used; I) The published and widely used RT-qPCR assay for the E-gene [4,5] combined with the SensiFAST TM Probe No-ROX One-Step Real-time PCR kit (Bioline®), and II) the commercial RealStar® SARS-CoV-2 RT-PCR kit 1.0 (Altona diagnostics, Hamburg, Germany). The RT-qPCR results (number of positives and Ct values) from the different approaches were compared to the RT-qPCR results from MagNA Pure purified samples.…”