Stability of beta-actin mRNA in plasma
N C Holford1, H S Sandhu, H Thakkar
1Department of Chemical Pathology, St. Thomas' Hospital, London, United Kingdom.
Annals of the New York Academy of Sciences
|October 8, 2008
Summary
Messenger RNA (mRNA) is not always stable in plasma. This study found that beta-actin mRNA levels significantly decreased within hours of blood collection, suggesting in vitro degradation.
Area of Science:
- Molecular Biology
- Biochemistry
- Clinical Diagnostics
Background:
- Messenger RNA (mRNA) is generally considered labile and susceptible to degradation.
- A prior study suggested that GAPDH mRNA in cell-free plasma could remain stable for up to 24 hours post-collection.
Purpose of the Study:
- To independently verify the stability of mRNA in cell-free plasma.
- To investigate the degradation kinetics of plasma beta-actin mRNA over time.
Main Methods:
- Blood samples were collected from a healthy volunteer in EDTA tubes and kept on ice.
- Plasma was isolated via double-centrifugation at 0, 1, 2, and 5 hours post-collection.
- RNA extraction, reverse transcription to cDNA, and quantitative PCR for beta-actin mRNA were performed.
Main Results:
- Plasma beta-actin mRNA levels significantly decreased from 0 hours to 2 hours (P=0.022).
- Further significant reductions in beta-actin mRNA were observed at 5 hours compared to 0 hours (P=0.004).
- Median beta-actin mRNA levels dropped from 0.12 at 0 hours to 0.012 at 2 hours and 0.0037 at 5 hours.
Conclusions:
- Plasma beta-actin mRNA levels decline over time after blood collection.
- The observed decrease in mRNA levels is likely due to degradation occurring in vitro after sample collection.
- These findings challenge the assumption of long-term mRNA stability in cell-free plasma samples.
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