Improved assay for fecal calprotectin.

Area of Science:

• Gastroenterology
• Clinical Chemistry
• Biomedical Engineering

Background:

• Fecal calprotectin is a key biomarker for lower gastrointestinal inflammatory and neoplastic diseases.
• Accurate and reproducible measurement of fecal calprotectin is crucial for effective disease diagnosis and management.
• Existing methods for fecal calprotectin measurement have limitations in yield and contamination risk.

Purpose of the Study:

• To develop and evaluate a novel fecal sample preparation procedure for enhanced fecal calprotectin measurement.
• To improve calprotectin yield, reduce contamination risk, and enhance the separation of high and low calprotectin levels.
• To assess the accuracy, reproducibility, and potential interferences of the new method.

Main Methods:

• A new fecal sample preparation procedure involving smaller sample size, higher dilution, dissociating agents, and closed disposable tubes was developed.
• Comparison with the original method [Roseth AG et al., 1992] focusing on calprotectin extraction yield and sample processing.
• Measurement of fecal calprotectin levels in healthy subjects and patients with pancolitis, and assessment of method reproducibility and interferences.

Main Results:

• The new method achieved a 78% extraction yield, a five-fold increase compared to the original method.
• Improved separation between high and low calprotectin levels was observed.
• Median calprotectin in healthy subjects was 26 microg/g, while pathological samples reached up to 30000 microg/g; measurements were accurate and reproducible with no significant interferences.

Conclusions:

• The novel fecal sample preparation procedure significantly enhances fecal calprotectin measurement.
• The improved method offers higher yield, better discrimination of disease states, and reliable results.
• This optimized procedure is suitable for accurate and reproducible fecal calprotectin quantification in clinical diagnostics.