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Standardizing leucocyte PNH clone detection: an international study.

Matthew Fletcher1, D Robert Sutherland, Liam Whitby

  • 1UK NEQAS for Leucocyte Immunophenotyping (UK NEQAS LI), Department of Haematology, Royal Hallamshire Hospital, Sheffield, United Kingdom.

Cytometry. Part B, Clinical Cytometry
|April 10, 2014
PubMed
Summary
This summary is machine-generated.

Standardized protocols improve paroxysmal nocturnal hemoglobinuria (PNH) detection accuracy. Stabilized PNH blood samples are suitable for high-sensitivity testing, reducing variation in PNH clone detection rates among experienced laboratories.

Keywords:
EQAparoxysmal nocturnal hemoglobinuriaquality assessmentquality controlstandardization

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Area of Science:

  • Hematology
  • Clinical Diagnostics
  • Flow Cytometry

Background:

  • Recent guidelines emphasize robust high-sensitivity detection of glycophosphatidylinositol-deficient cells in paroxysmal nocturnal hemoglobinuria (PNH).
  • Accurate PNH diagnosis relies on detecting PNH cell populations on red blood cells and white blood cells.

Purpose of the Study:

  • To evaluate the suitability of stabilized whole PNH blood samples for high-sensitivity detection of PNH clones.
  • To compare the performance of standardized protocols versus in-house methods for PNH testing.

Main Methods:

  • UK NEQAS LI distributed stabilized PNH blood samples (normal, 0.1% PNH, 8% PNH) with standardized reagents and SOPs to 19 international laboratories.
  • Laboratories tested samples using both standardized and their own in-house protocols.
  • Results were compared between standardized, in-house, and full External Quality Assessment (EQA) groups.

Main Results:

  • No difference in PNH clone detection rates between standardized and in-house methods.
  • Standardized approach showed lower variation around the median (e.g., IQR of 0.48% vs. 1.29% for 8% PNH neutrophils).
  • Full EQA group exhibited the greatest variation (IQR 1.7%), significantly different from the standardized cohort (P<0.001).

Conclusions:

  • Stabilized whole PNH blood samples are appropriate for high-sensitivity testing with recommended reagent cocktails and protocols.
  • Careful selection of conjugates alongside standardized protocols is crucial for accurate PNH detection.
  • Standardized approaches reduce inter-laboratory variation in PNH clone detection, even among experienced labs.