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A novel multiplex polymerase chain reaction assay for profile analyses of gene expression in peripheral blood

Xingwang Jia1, Haiyue Ju, Li Yang

  • 1Department of Clinical Biochemistry, State Key Laboratory of Kidney Disease, Chinese PLA General Hospital, 100853 Beijing, China.

BMC Cardiovascular Disorders
|July 12, 2012
PubMed

Insights

A new multiplex PCR method accurately measures gene expression in blood for coronary artery disease (CAD) risk assessment. Specific gene expression profiles correlate with CAD, offering potential diagnostic insights.

Area of Science:

  • Molecular Biology
  • Cardiovascular Disease Research
  • Biomarker Discovery

Background:

  • Inflammation plays a critical role in atherosclerosis pathogenesis, driven by aberrant cytokine gene expression.
  • Accurate methods for observing whole blood gene expression are needed for diagnosing coronary artery disease (CAD) and related conditions.

Purpose of the Study:

  • To develop and validate a precise multiplex polymerase chain reaction (multi-PCR) method for simultaneous measurement of multiple CAD-related genes.
  • To explore the relationship between gene expression profiles, clinical parameters (glucose, lipids), and atherosclerosis status in a human cohort.
  • To establish a gene expression analysis system for potential CAD risk assessment.

Main Methods:

  • A novel multi-PCR assay was developed to measure 15 CAD-related genes and 2 housekeeping genes in peripheral blood.
  • Assay precision was evaluated, and a diagnostic procedure was developed using data from 148 subjects categorized by atherosclerosis presence and type (calcified vs. non-calcified plaques).
  • Gene expression, glucose, lipid levels, and clinical data were analyzed to identify correlations.

Main Results:

  • The multi-PCR system demonstrated good precision with acceptable within-run and between-run coefficients of variation.
  • Positive correlations were observed between glucose levels and MCSF, HMOX1, or TNFalpha gene expression.
  • Significantly elevated expression of IL1B, IL6, IL8, and MCP-1 was found in patients with non-calcified plaques compared to controls.

Conclusions:

  • A novel multiplex gene expression analysis system for whole blood has been successfully developed and validated.
  • Gene expression patterns in peripheral blood are demonstrably related to the presence and type of coronary artery disease.
  • This system holds promise as a tool for the risk assessment of cardiovascular diseases (CVDs) and associated conditions.
Abstract

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