Paroxysmal nocturnal hemoglobinuria testing by flow cytometry. Evaluation of the REDQUANT and CELLQUANT kits

Am J Clin Pathol. 2000 Nov;114(5):798-806. doi: 10.1309/GFCJ-NXLC-BRXB-YUJ4.

Abstract

The commercially available REDQUANT and CELLQUANT kits were evaluated in a clinical flow cytometry laboratory using healthy volunteers and a small series of 5 patients with paroxysmal nocturnal hemoglobinuria (PNH). Most samples also were tested with a "home brew" antibody panel for CD66b expression on granulocytes and CD14 expression on monocytes. The REDQUANT and CELLQUANT kits in tandem correctly identified all 5 PNH samples with no false-positive results. The 1 PNH sample from a patient with a negative Ham test result also was identified correctly. One marker, CD59 on granulocytes, when considered alone, had a high false-positive rate, and I suggest lowering the fluorescence intensity criteria for identifying decreased antigen expression. The home-brew tube also performed well, identifying all 5 patients, and may serve as a supplement in difficult specimens. The evaluated kits performed well. However, the validation series was small, which is likely to be a problem at any single institution. The provision of a reproducible manufacturer-defined expression level for normal cells adds another degree of confidence that the test correctly identifies patients with this potentially life-threatening disorder while avoiding false-positive results.

MeSH terms

  • Adult
  • Aged
  • Antigens, CD
  • Antigens, Neoplasm*
  • CD55 Antigens / analysis
  • CD59 Antigens / analysis
  • Cell Adhesion Molecules*
  • Erythrocytes / immunology
  • Evaluation Studies as Topic
  • Female
  • Flow Cytometry*
  • Fluorescein-5-isothiocyanate
  • Fluorescent Dyes
  • GPI-Linked Proteins
  • Granulocytes / immunology
  • Hemoglobinuria, Paroxysmal / blood*
  • Humans
  • Immunophenotyping
  • Lipopolysaccharide Receptors / analysis
  • Male
  • Membrane Glycoproteins / analysis
  • Middle Aged
  • Monocytes / immunology
  • Reagent Kits, Diagnostic*

Substances

  • Antigens, CD
  • Antigens, Neoplasm
  • CD55 Antigens
  • CD59 Antigens
  • CEACAM8 protein, human
  • Cell Adhesion Molecules
  • Fluorescent Dyes
  • GPI-Linked Proteins
  • Lipopolysaccharide Receptors
  • Membrane Glycoproteins
  • Reagent Kits, Diagnostic
  • Fluorescein-5-isothiocyanate