RT Journal Article
T1 Laboratory protocol for the digital multiplexed gene expression analysis of nasopharyngeal swab samples using the NanoString nCounter system.
A1 Garcia-Aranda, Marilina
A1 Lopez-Rodriguez, Inmaculada
A1 Garcia-Gutierrez, Susana
A1 Padilla-Ruiz, Maria
A1 de-Luque, Vanessa
A1 Hortas, Maria Luisa
A1 Diaz, Tatiana
A1 Alvarez, Martina
A1 Barragan-Mallofret, Isabel
A1 Redondo, Maximino
K1 Digital RNA quantification
K1 Immunology
K1 Gene expression
K1 Nasopharyngeal swab
K1 Respiratory infection
AB This paper describes a laboratory protocol to perform the NanoString nCounter Gene Expression Assay from nasopharyngeal swab samples.  It is urgently necessary to identify factors related to severe symptoms of respiratory infectious diseases, such as COVID-19, in order to assess the possibility of establishing preventive or preliminary therapeutic measures and to plan the services to be provided on hospital admission. At present, the samples recommended for microbiological diagnosis are those taken from the upper and/or the lower respiratory tract.  The NanoString nCounter Gene Expression Assay is a method based on the direct digital detection of mRNA molecules by means of target-specific, colour-coded probe pairs, without the need for mRNA conversion to cDNA by reverse transcription or the amplification of the resulting cDNA by PCR. This platform includes advanced analysis tools that reduce the need for bioinformatics support and also offers reliable sensitivity, reproducibility, technical robustness and utility for clinical application, even in RNA samples of low RNA quality or concentration, such as paraffin-embedded samples. Although the protocols for the analysis of blood or formalin-fixed paraffin-embedded samples are provided by the manufacturer, no corresponding protocol for the analysis of nasopharyngeal swab samples has yet been established. Therefore, the approach we describe could be adopted to determine the expression of target genes in samples obtained from nasopharyngeal swabs using the nCOUNTER technology.
PB Faculty of 1000
YR 2022
FD 2022-09-30
LK http://hdl.handle.net/10668/21681
UL http://hdl.handle.net/10668/21681
LA en
NO García Aranda M, López-Rodríguez I, García-Gutiérrez S, Padilla-Ruiz M, de Luque V, Hortas ML, et al. Laboratory protocol for the digital multiplexed gene expression analysis of nasopharyngeal swab samples using the NanoString nCounter system. F1000Res. 2022 Feb 2;11:133
NO This research was supported by grants from Instituto de Salud Carlos III through the project REDISSEC COV20/0059 awarded to MR; Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía through the projects CV207697_COVID to MR and CV20-62050_COVID to IB, as well as by donations from Fundación Fuerte (Marbella, Spain) and Ms Isabel María Luque Bardón to the Research and Innovation Unit at Hospital Costa del Sol.
DS RISalud
RD Apr 17, 2025