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Preanalytical classical and alternative complement pathway activity loss.

Koen O A Vercauteren1, Stijn Lambrecht1, Joris Delanghe1

  • 1Department of clinical chemistry, Ghent University Hospital, Belgium.

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|August 6, 2019
PubMed
Summary
This summary is machine-generated.

Complement functional assays (CH50 and AP50) are stable for one week on ice. Serum at room temperature is suitable for 24 hours, while EDTA plasma offers extended stability for alternative pathway analysis up to four days.

Keywords:
alternative and classical complement pathwaycomplement C3complement C3dcomplement haemolytic activity assaypreanalytical phase

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Area of Science:

  • Immunology
  • Clinical Chemistry

Background:

  • Complement functional analyses assess the integrity of the complement cascade and are crucial for diagnosing complement deficiencies.
  • Preanalytical factors like time, temperature, and sample type can impact complement component stability, leading to inaccurate test results.
  • Understanding preanalytical stability is essential for reliable complement testing.

Purpose of the Study:

  • To evaluate the preanalytical stability of classical (CH50) and alternative pathway (AP50) functional assays.
  • To determine the influence of time, temperature, and EDTA on complement component stability in different sample types.

Main Methods:

  • Assessed CH50 and AP50 functional assay stability under various conditions (time, temperature, EDTA).
  • Utilized nephelometry (C3d) and immunofixation (C3c) for supporting investigations.
  • Employed hemolytic activity assays and C5b-9 ELISA for quantitative pathway function determination.

Main Results:

  • CH50 and AP50 demonstrated stability for approximately one week in serum samples stored on ice.
  • CH50 activity declined twice as rapidly in EDTA plasma compared to serum at room temperature.
  • AP50 activity decreased twice as slowly in EDTA plasma versus serum at room temperature.

Conclusions:

  • Serum on ice is the preferred specimen for functional complement analyses.
  • Serum at room temperature (up to 24 hours) is acceptable for both classical and alternative pathway analyses.
  • EDTA plasma is suitable for alternative pathway analyses for up to four days due to C3 stabilization, retaining at least 85% of baseline activity.