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Related Experiment Videos

Solid-phase competitive luminescence immunoassay for lysozyme in faeces

P Gao1, M R John, H Schmidt-Gayk

  • 1Department of Endocrinology and Oncology, Laboratory Group, Heidelberg, Germany.

Clinica Chimica Acta; International Journal of Clinical Chemistry
|August 14, 1995
PubMed
Summary

A new luminescence immunoassay (LIA) accurately measures faecal lysozyme, aiding in inflammatory bowel disease diagnosis. This simple assay differentiates healthy individuals from those with Crohn's disease or ulcerative colitis.

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Area of Science:

  • Biochemistry
  • Immunology
  • Clinical Diagnostics

Background:

  • Faecal lysozyme is an indicator of gastrointestinal inflammation.
  • Accurate measurement of faecal lysozyme is crucial for diagnosing and monitoring inflammatory bowel diseases (IBD).

Purpose of the Study:

  • To develop and validate a simple, solid-phase luminescence immunoassay (LIA) for quantifying faecal lysozyme.
  • To assess the utility of this assay in differentiating between healthy individuals and patients with IBD.

Main Methods:

  • A solid-phase luminescence immunoassay (LIA) was developed using polystyrene beads coated with a polyclonal capture antibody and acridinium ester-labelled human lysozyme tracer.
  • Samples were incubated overnight, followed by washing and measurement of emitted light using an automated luminometer.

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  • Assay validation included sensitivity, dilution recovery, and intra- and inter-assay coefficient of variation (CV) assessments.

    • Main Results:

    • The assay demonstrated a sensitivity of 0.02 mg/l with good dilution recoveries (88-108%).
    • Intra- and inter-assay CVs were within acceptable ranges, indicating assay reproducibility.
    • Faecal lysozyme levels were significantly elevated in patients with Crohn's disease (median 1.75 mg/l) and ulcerative colitis (median 1.11 mg/l) compared to healthy controls (median 0.28 mg/l).

    Conclusions:

    • The developed luminescence immunoassay (LIA) is a simple, sensitive, and reproducible method for determining faecal lysozyme.
    • Elevated faecal lysozyme levels effectively distinguish individuals with inflammatory bowel disease (IBD) from healthy subjects.
    • This assay represents a valuable tool for the diagnosis and evaluation of inflammatory bowel disease.