Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples of Sulfur Mustard-Exposed Veterans

Objective: in spite of accumulating information about pathological aspects of sulfurmustard (sm), the precise mechanism responsible for its effects is not well understood.circulating micrornas (mirnas) are promising biomarkers for disease diagnosis andprognosis. accurate normalization using appropriate reference genes, is a critical step inmirna expression studies. in this study, we aimed to identify appropriate reference genefor microrna quantification in serum samples of sm victims.materials and methods: in this case and control experimental study, using quantitativereal-time polymerase chain reaction (qrt-pcr), we evaluated the suitability of a panelof small rnas including snord38b, snord49a, u6, 5s rrna, mir-423-3p, mir-191,mir-16 and mir-103 in sera of 28 sm-exposed veterans of iran-iraq war (1980-1988)and 15 matched control volunteers. different statistical algorithms including genorm,norm?nder, best-keeper and comparative delta-quantification cycle (cq) method wereemployed to find the least variable reference gene.results: mir-423-3p was identified as the most stably expressed reference gene, and mir-103 and mir-16 ranked after that.conclusion: we demonstrate that non-mirna reference genes have the least stabilityin serum samples and that some house-keeping mirnas may be used as morereliable reference genes for mirnas in serum. in addition, using the geometric meanof two reference genes could increase the reliability of the normalizers.