| Autor: |
Borowitz, Michael J., Craig, Fiona E., DiGiuseppe, Joseph A., Illingworth, Andrea J., Rosse, Wendell, Sutherland, D. Robert, Wittwer, Carl T., Richards, Stephen J. |
| Zdroj: |
Cytometry Part B: Clinical Cytometry; July 2010, Vol. 78 Issue: 4 p211-230, 20p |
| Abstrakt: |
Background:Paroxysmal nocturnal hemoglobinuria PNH is a rare hematopoietic stem cell disorder characterized by a somatic mutation in the PIGAgene, leading to a deficiency of proteins linked to the cell membrane via glycophosphatidylinositol GPI anchors. While flow cytometry is the method of choice for identifying cells deficient in GPIlinked proteins and is, therefore, necessary for the diagnosis of PNH, to date there has not been an attempt to standardize the methodology used to identify these cells.Methods:In this document, we present a consensus effort that describes flow cytometric procedures for detecting PNH cells.Results:We discuss clinical indications and offer recommendations on data interpretation and reporting but mostly focus on analytical procedures important for analysis. We distinguish between routine analysis defined as identifying an abnormal population of 1 or more and highsensitivity analysis in which as few as 0.01 PNH cells are detected. Antibody panels and gating strategies necessary for both procedures are presented in detail. We discuss methods for assessing PNH populations in both white blood cells and red blood cells and the relative advantages of measuring each. We present steps needed to validate the more elaborate highsensitivity techniques, including the need for careful titration of reagents and determination of background rates in normal populations, and discuss technical pitfalls that might affect interpretation.Conclusions:This document should both enable laboratories interested in beginning PNH testing to establish a valid procedure and allow experienced laboratories to improve their techniques. © 2010 Clinical Cytometry Society |
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