QuantiNova Multiplex RT-PCR Kit (2500)

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Publication protocol

Nucleic acids were pooled from multiple extractions of one SARS-CoV-2-positive stool sample using the QIAsymphony DSP Virus/Pathogen Kit (Qiagen, Hilden, Germany). This stool sample was from a 5-year-old child with COVID-19 [7] and was chosen because of high initial PCR signals and sufficient sample availability to generate large quantities of eluate for further distribution. Of note, no PCR inhibition was observed for detection of the spiked-in extraction RNA control (QuantiNova IC Probe Assays Red 650, Qiagen). A 1:10 dilution series was prepared and aliquots were labelled in a blinded fashion to be shipped on dry ice to participating laboratories in March 2020. Participants were instructed to perform the diagnostic assays used at their centre for SARS-CoV-2 detection in quadruplicate using 5 µL of the aliquot per reaction. All results were reported back to the initiating laboratory (Laboratory 1) before the results were unblinded. The details of all these PCR-based assays are summarised in the Table.

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