Nucleosomes in Serum as a Marker for Cell Death
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Stefan Holdenrieder
Abstract
The concentration of nucleosomes is elevated in blood of patients with diseases which are associated with enhanced cell death. In order to detect these circulating nucleosomes, we used the Cell Death Detection-ELISAplus (CDDE) from Roche Diagnostics (Mannheim, Germany) (details at http:\\biochem.roche.com). For its application in liquid materials we performed various modifications: we introduced a standard curve with nucleosome-rich material, which enabled direct quantification and improved comparability of the values within (CVintraassay:3.0–4.1%) and between several runs (CVinterassay:8.6–13.5%), and tested the analytical specificity of the ELISA.
Because of the fast elimination of nucleosomes from circulation and their limited stability, we compared plasma and serum matrix and investigated in detail the pre-analytical handling of serum samples which can considerably influence the test results. Careless venipuncture producing hemolysis, delayed centrifugation and bacterial contamination of the blood samples led to false-positive results; delayed stabilization with EDTA and insufficient storage conditions resulted in false-negative values. At temperatures of −20 °C, serum samples which were treated with 10 mM EDTA were stable for at least 6 months. In order to avoid possible interfering factors, we recommend a schedule for the pre-analytical handling of the samples.
As the first stage, the possible clinical application was investigated in the sera of 310 persons. Patients with solid tumors (n=220; mean=361 Arbitrary Units (AU)) had considerably higher values than healthy persons (n=50; mean=30 AU; p=0.0001) and patients with inflammatory diseases (n=40; mean= 296 AU; p=0.096). Within the group of patients with tumors, those in advanced stages (UICC 4) showed significantly higher values than those in early stages (UICC 1–3) (p=0.0004).
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
- The Impact of Cardiac Natriuretic Peptide Determination on the Diagnosis and Management of Heart Failure
- Models for Combining Random and Systematic Errors. Assumptions and Consequences for different Models
- Nucleosomes in Serum as a Marker for Cell Death
- Comparative Nuclear Magnetic Resonance Studies of Water Permeability of Red Blood Cells from Maternal Venous Blood and Newborn Umbilical Cord Blood
- Optimization of Single-Stranded Conformation Polymorphism (SSCP) Analysis for Screening for the Estrogen Receptor-α Gene Polymorphism P325P
- Serum Total Renin after Tubal Sterilization
- Oxidative Stress and Male IGF-1, Gonadotropin and Related Hormones in Diabetic Patients
- New PCR-Based Method for the Sp1 Site Polymorphism in the COL1A1 Gene
- Comparability of Serum and Plasma Concentrations of Haemostasis Activation Markers
- Importance of Validation of Immunoassays for Intact Proinsulin
- Plant Sterol-Enriched Margarine Lowers Plasma LDL in Hyperlipidemic Subjects with low Cholesterol Intake: Effect of Fibrate Treatment
- Menopause, Coronary Artery Disease and Antioxidants
- Arterial Tissue of Arsenic, Selenium and Iron in Blackfoot Disease Patients
- Evaluation of Aution Max AX-4280 Automated Urine Test-Strip Analyser
- Evaluation of Automated Enzyme Immunoassays for the Detection of Antibodies to Extractable Nuclear Antigens
- Metrological Requirements for Clinical Laboratories
