Abstract
Thispaper describes alaboratory protocol to perform theNanoStringnCounterGene Expression Assay from nasopharyngeal swabsamples. It is urgently necessary toidentifyfactors relatedtosevere symptoms of respiratory infectious diseases,such asCOVID-19,in ordertoassess the possibility ofestablishingpreventive orpreliminarytherapeutic measuresand to planthe services to be providedonhospital admission.At present,the samples recommended for microbiological diagnosis are thosetakenfromthe upperand/or the lowerrespiratory tract. TheNanoStringnCounterGene Expression Assay is a methodbasedonthedirectdigitaldetection of mRNA moleculesby means oftarget-specific, colour-coded probe pairs, withoutthe needformRNA conversion to cDNA by reverse transcription or the amplification of the resulting cDNA byPCR. This platform includesadvanced analysis tools that reduce the need for bioinformatics supportand alsooffersreliable sensitivity, reproducibility, technicalrobustnessand utility for clinical application, even inRNAsamples of lowRNAquality or concentration,such as paraffin-embedded samples.Although the protocols for the analysis of blood or formalin-fixed paraffin-embedded samples areprovidedby the manufacturer,no correspondingprotocol for the analysis of nasopharyngeal swab samples hasyetbeenestablished.Therefore,the approach we describe could be adopted todeterminethe expression of target genes insamples obtained fromnasopharyngeal swabsusingthenCOUNTERtechnology.
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