Preanalytical, analytical and postanalytical considerations in circulating microRNAs measurement

Abstract

Highlights • Methods used for miRNAs expression profiling are quantitative reverse transcription polymerase chain reaction, microarrays, next generation sequencing and droplet digital PCR • To obtain reproducible and accurate miRNAs expression profiling detection, it is crucial to strictly standardize the entire process, starting from choosing the specimen type until the normalization strategy on the interpretation of miRNAs expression profiling detection •Due to the critical impact of the normalization strategy on the miRNAs expression, the choice of normalization agent is of great importance Microribonucleic acids (miRNAs) have emerged as a new category of biomarkers for many human diseases like cancer, cardiovascular and neurodegenerative disorders. MicroRNAs can be detected in various body fluids including blood, urine and cerebrospinal fluid. However, the literature contains conflicting results for circulating miRNAs, which is the main barrier to using miRNAs as non-invasive biomarkers. This variability in results is largely due to differences between studies in sample processing methodology, miRNA quantification and result normalization. The purpose of this review is to describe the various preanalytical, analytical and postanalytical factors that can impact miRNA detection accuracy and to propose recommendations for the standardization of circulating miRNAs measurement.