PERFORMANCE CHARACTERISTICS OF CONSENSUAL APPROACHES FOR SMALL AND MINOR PAROXYSMAL NOCTURNAL HEMOGLOBINURIA CLONE DETERMINATION BY FLOW CYTOMETRY

Konference: 2013 18th Congress of the European Hematology Association - účast ČR

Kategorie: Maligní lymfomy a leukémie

Téma: Myelodysplastic syndromes and bone marrow failure syndromes incl. PNH - Biology

Číslo abstraktu: B1430

Autoři: MUDr. Iuri Marinov, CSc.; Martina Kohoutová; Mgr. Vlasta Tkáčová; Adam Pešek; Doc.MUDr. Jaroslav Čermák, CSc.; doc. MUDr. Petr Cetkovský, Ph.D.

Background:

Comparison of individual approaches for paroxysmal nocturnal hemoglobinuria (PNH) clones  evaluation by flow cytometry (FCM) as described in the 2010 international clinical cytometry society (ICCS) guidelines has not been reported in literature.

Aims:

To rule out the influence of different consensual approaches and strategies for small and minor PNH clone evaluation by flow cytometry on final results, thus leading to consistant interlaboratory variabilty

Methods:

We analyzed the performance characteristics of  4, 5 and 6 color protocols for white blood cell (WBC), one and two color protocols for red blood cell (RBC) evaluation for different target PNH clones and compared results from routine PNH patient analysis.

Results:

Coefficient of variation (CV) for precision / reproducibility analysis ranged from 0.67 % / 1.49 % to 2.56 % / 3.09 % for granulocytes, from 0.93 % / 3.09 % to 7.76 % / 12.06 % for monocytes and from 0.41 % / 4.73 % to 6.53 % / 5.1 % for RBCs. Linear regression analysis revealed excellent correlation (r = 0.99 %), Wilcoxon ranks test showed no statistically significant differences (P > 0.05), Bland-Altman analysis demonstrated performance agreement with mean bias ranging from 0.02 to 2.2.

Summary / Conclusion:

Our results confirmed very good performance characteristics for precision and reproducibility analysis, excellent correlation and favorable agreement between protocols, underlining the crucial role of optimally selected glycophosphatidylinositol (GPI)-specific reagents and appropriate conjugates and secondary role of the number and type of gating reagents, tubes per test and corresponding gating strategy.  With respect to this, reported high interlaboratory variability is considerably related to incorrect performance and /or insufficient experience with PNH testing by flow cytometry.

Keywords: Flow cytometry, PNH

Abstrakta v časopise Haematologica 2013, Suppl1

Online Program

Datum přednesení příspěvku: 15. 6. 2013