Options
High Throughput qPCR Expression Profiling of Circulating MicroRNAs Reveals Minimal Sex and Sample Timing-Related Variation in Plasma of Healthy Volunteers
Author(s)
Editor(s)
Date Issued
2015-12-23
Date Available
2019-04-23T09:22:52Z
Abstract
MicroRNAs are a class of small non-coding RNA that regulate gene expression at a posttranscriptional level. MicroRNAs have been identified in various body fluids under normal conditions and their stability as well as their dysregulation in disease opens up a new field for biomarker study. However, diurnal and day-to-day variation in plasma microRNA levels, and differential regulation between males and females, may affect biomarker stability. A QuantStudio 12K Flex Real-Time PCR System was used to profile plasma microRNA levels using OpenArray in male and female healthy volunteers, in the morning and afternoon, and at four time points over a one month period. Using this system we were able to run four OpenArray plates in a single run, the equivalent of 32 traditional 384-well qPCR plates or 12,000 data points. Up to 754 microRNAs can be identified in a single plasma sample in under two hours. 108 individual microRNAs were identified in at least 80% of all our samples which compares favourably with other reports of microRNA profiles in serum or plasma in healthy adults. Many of these microRNAs, including miR-16-5p, miR-17-5p, miR-19a-3p, miR-24-3p, miR-30c-5p, miR-191-5p, miR-223-3p and miR-451a are highly expressed and consistent with previous studies using other platforms. Overall, microRNA levels were very consistent between individuals, males and females, and time points and we did not detect significant differences in levels of microRNAs. These results suggest the suitability of this platform for microRNA profiling and biomarker discovery and suggest minimal confounding influence of sex or sample timing. However, the platform has not been subjected to rigorous validation which must be demonstrated in future biomarker studies where large differences may exist between disease and control samples.
Sponsorship
European Commission - Seventh Framework Programme (FP7)
Science Foundation Ireland
Other Sponsorship
Iraqi Ministry of Higher Education and Scientific Research
Type of Material
Journal Article
Publisher
PLoS
Journal
PLoS ONE
Volume
10
Issue
12
Start Page
1
End Page
20
Copyright (Published Version)
2015 the Authors
Language
English
Status of Item
Peer reviewed
ISSN
1932-6203
This item is made available under a Creative Commons License
File(s)
No Thumbnail Available
Name
High Throughput qPCR Expression Profiling of Circulating MicroRNAs Reveals Minimal Sex- and Sample Timing-Related Variation in Plasma of Healthy Volunteers.pdf
Size
1.32 MB
Format
Adobe PDF
Checksum (MD5)
d736d749f3f32adbe54702bff763e01d
Owning collection