Abstract
BackgroundMost studies of microRNA (miRNA) and disease have examined tissue-specific expression in limited numbers of samples. The presence of circulating miRNAs in plasma samples provides the opportunity to examine prospective associations between miRNA expression and disease in initially healthy individuals. However, little data exist on the reproducibility of miRNAs in stored plasma.MethodsWe used Real-Time PCR to measure 61 pre-selected microRNA candidates in stored plasma. Coefficients of variation (CVs) were used to assess inter-assay reliability (n = 15) and within-person stability over one year (n = 80). Intraclass correlation coefficients (ICCs) and polychoric correlation coefficients were used to assess within-person stability and delayed processing reproducibility (whole blood stored at 4°C for 0, 24 and 48 hours; n = 12 samples).ResultsOf 61 selected miRNAs, 23 were detected in at least 50% of samples and had average CVs below 20% for inter-assay reproducibility and 31 for delayed processing reproducibility. Ten miRNAs were detected in at least 50% of samples, had average CVs below 20% and had ICCs above 0.4 for within-person stability over 1–2 years, six of which satisfied criteria for both interassay reproducibility and short-term within-person stability (miR-17-5p, -191-5p, -26a-5p, -27b-3p, -320a, and -375) and two all three types of reproducibility (miR-27b-3p and -26a-5p). However, many miRNAs with acceptable average CVs had high maximum CVs, most had low expression levels, and several had low ICCs with delayed processing.ConclusionsAbout a tenth of miRNAs plausibly related to chronic disease were reliably detected in stored samples of healthy adults.
Highlights
Micro RNAs have recently emerged as key post-transcriptional regulators of gene expression
Of 61 selected Micro RNAs (miRNAs), 23 were detected in at least 50% of samples and had average coefficient of variation (CV) below 20% for inter-assay reproducibility and 31 for delayed processing reproducibility
Ten miRNAs were detected in at least 50% of samples, had average CVs below 20% and had Intraclass correlation coefficients (ICCs) above 0.4 for within-person stability over 1–2 years, six of which satisfied criteria for both interassay reproducibility and short-term within-person stability and two all three types of reproducibility
Summary
Micro RNAs (miRNAs) have recently emerged as key post-transcriptional regulators of gene expression. MiRNAs are ~22bp single-stranded RNA segments that silence gene expression by binding to complementary messenger RNA (mRNA). This binding represses translation and speeds mRNA degradation [1]. The number of promising studies examining miRNAs related to CVD and cancer has grown substantially in recent years, most studies in humans have been cross-sectional and generally limited to 300 or fewer subjects [2]. Prospective cohorts with long-term stored plasma samples provide a unique opportunity to economically and efficiently study the prospective relationship between circulating miRNA expression and chronic disease. Most studies of microRNA (miRNA) and disease have examined tissue-specific expression in limited numbers of samples. Little data exist on the reproducibility of miRNAs in stored plasma
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