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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
PubMed

Insights

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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