Validation of reference gene stability for miRNA quantification by reverse transcription quantitative PCR in the peripheral blood of patients with COVID-19 critical illness

• DOI: 10.1371/journal.pone.0286871
• Publication Date: 2023-08-29T17:46:04Z
• Authors: Amanda Formosa; Erica Acton; Amy Lee; Paul Turgeon; Shehla Izhar; Pamela Plant; Jim N. Tsoporis; Sabri Soussi; Uriel Trahtemberg; Andrew Baker; Claudia C. dos Sa...
• Keywords: Reference Genes

The COVID-19 pandemic has created an urgency to study the host gene response that leads variable clinical presentations of disease, particularly critical illness response. miRNAs have been implicated in mechanism immune dysregulation and thus hold potential as biomarkers and/or therapeutic agents with application. Hence, further analyses their altered expression is warranted. An important basis for this identifying appropriate reference genes high quality analysis studies. In current report, NanoString technology was used 798 peripheral blood 24 critically ill patients, 12 had were negative. A list potentially stable candidate generated included ten miRNAs. top six analyzed using reverse transcription quantitative polymerase chain reaction (RT-qPCR) a total 41 patients so apply standard computational algorithms validating genes, namely geNorm, NormFinder, BestKeeper RefFinder. There general agreement among all four ranking miRNAs: miR-186-5p, miR-148b-3p, miR-194-5p miR-448. detailed output rankings led conclusion miR-186-5p miR-148b-3p are miRNA studies PaxGene tubes disease.