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Related Experiment Video

Updated: Jun 29, 2026

Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays
13:58

Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays

Published on: September 26, 2011

Normalization of circulating nucleic acid results.

N C Holford1, A Ramoutar, A N Butt

  • 1Department of Chemical Pathology, Guy's and St. Thomas' Hospital NHS Trust, London, United Kingdom.

Annals of the New York Academy of Sciences
|October 8, 2008
PubMed
Summary
This summary is machine-generated.

Beta-actin messenger RNA (mRNA) levels are elevated in hyperthyroidism, impacting its use as a reference gene. Researchers found normalizing gene expression to beta-actin may not be suitable in all clinical studies.

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Published on: June 30, 2018

Area of Science:

  • Molecular Biology
  • Endocrinology
  • Clinical Chemistry

Background:

  • Circulating nucleic acids (DNA and mRNA) have diagnostic applications in fetal medicine, oncology, and transplantation.
  • Messenger RNA (mRNA) quantification in blood often uses reference genes, like beta-actin mRNA, for normalization.
  • Thyroid hormone status can influence gene expression, necessitating investigation into suitable reference genes.

Purpose of the Study:

  • To investigate the impact of thyroid hormone status on the expression of thyroid hormone-dependent genes.
  • To evaluate the suitability of beta-actin mRNA as a reference gene in hyperthyroid subjects.

Main Methods:

  • Blood samples were collected from healthy subjects and patients with hyperthyroidism.
  • Messenger RNA (mRNA) was extracted, and complementary DNA (cDNA) was synthesized.
  • Quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to measure Na/K-ATPase, T3 receptor, and beta-actin cDNA.

Main Results:

  • Median beta-actin mRNA levels were significantly higher in hyperthyroid subjects compared to healthy controls (P < 0.00042).
  • T3 receptor mRNA normalized to beta-actin was significantly higher in hyperthyroidism (P < 0.05).
  • However, T3 receptor mRNA normalized to total RNA was significantly lower in hyperthyroidism (P < 0.00015).

Conclusions:

  • Beta-actin mRNA expression is significantly affected by thyroid hormone status.
  • Normalizing gene expression results to beta-actin mRNA may not be appropriate in conditions like hyperthyroidism.
  • Alternative normalization strategies, such as using total RNA, may be necessary for accurate gene expression analysis in certain clinical contexts.