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Published on: October 26, 2017
Reproducibility of Circulating MicroRNAs in Stored Plasma Samples
Monica L Bertoia1, Kimberly A Bertrand2, Sherilyn J Sawyer3
1Department of Nutrition, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, United States of America.
Background:
Most 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.
Methods:
We 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).
Results:
Of 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.
Conclusions:
About a tenth of miRNAs plausibly related to chronic disease were reliably detected in stored samples of healthy adults.
Insights
Reproducibility of microRNAs (miRNAs) in stored plasma is crucial for disease research. This study found that about 10% of candidate miRNAs showed reliable detection in healthy adults, supporting their use in future studies.
Area of Science:
- Biomarker discovery
- Molecular diagnostics
- Genomics and transcriptomics
Background:
- Limited studies assess microRNA (miRNA) expression in disease using tissue samples.
- Circulating miRNAs in plasma offer potential for prospective disease association studies in healthy individuals.
- Reproducibility data for miRNAs in stored plasma are scarce.
Purpose of the Study:
- To evaluate the reproducibility of detecting microRNAs (miRNAs) in stored plasma samples.
- To assess the reliability of specific miRNA candidates for use in large-scale epidemiological studies.
Main Methods:
- Real-Time PCR was used to quantify 61 pre-selected microRNA candidates in stored plasma.
- Inter-assay reliability, within-person stability over time, and delayed processing reproducibility were assessed using coefficients of variation (CVs) and intraclass correlation coefficients (ICCs).
Main Results:
- Twenty-three of 61 miRNAs met criteria for inter-assay reproducibility and delayed processing.
- Ten miRNAs demonstrated within-person stability over 1-2 years.
- Two miRNAs (miR-27b-3p and miR-26a-5p) showed reproducibility across all assessed conditions.
Conclusions:
- Approximately 10% of candidate microRNAs (miRNAs) related to chronic disease can be reliably detected in stored plasma from healthy adults.
- These findings support the potential utility of specific circulating miRNAs as biomarkers in stored samples.
- Further validation is needed, as many miRNAs exhibited variability or low expression levels.
