Automated analysis of GPI-deficient leukocyte flow cytometric data using GemStone™

Cytometry. Part B, Clinical Cytometry
David T MillerC Bruce Bagwell

Abstract

Flow Cytometry is the standard for the detection of glycosylphosphatidylinositol (GPI)-deficient clones in paroxysmal nocturnal hemoglobinuria (PNH) and related disorders. Although the International Clinical Cytometry Society (ICCS) and the International PNH Interest Group (IPIG) have published guidelines for PNH assays, data analysis has not been standardized. Current analyses use manual gating to enumerate PNH cells. We evaluate an automated approach to identify GPI-deficient leukocytes using a GemStone™ (Verity Software House) probability state model (PSM). Five hundred and thirty patient samples were assayed on BD Canto II flow cytometers using a stain-lyse-wash technique. Populations were defined using CD15, CD45, CD64 and side scatter. GPI-deficient myeloid cells were recognized as FLAER-, CD24-, and dim or absent CD16. GPI-deficient monocytic cells were identified as FLAER- and CD14-. The data were not censored in any way. A PSM was designed to enumerate monocytic and myeloid cells by adjusting the peaks and line spreads of the data, and recording the normal and GPI-deficient counts. No operator adjustments were made to the automated analysis. By human analysis, 53 of 530 samples showed GPI-deficient clones. Automated an...Continue Reading

References

Jul 30, 2005·Blood·Charles ParkerUNKNOWN International PNH Interest Group
Jul 31, 2007·Clinics in Laboratory Medicine·Stephen J Richards, David Barnett
Apr 3, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Michael J Boedigheimer, John Ferbas
Jan 13, 2009·Immunology·Pratip K ChattopadhyayMario Roederer
Feb 21, 2009·American Journal of Clinical Pathology·William G Finn
Dec 4, 2009·Advances in Bioinformatics·Wade T Rogers, Herbert A Holyst
Jun 10, 2010·Cytometry. Part B, Clinical Cytometry·Michael J BorowitzUNKNOWN Clinical Cytometry Society
Dec 1, 2010·Methods in Molecular Biology·C Bruce Bagwell

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Citations

Mar 14, 2013·Biochimica Et Biophysica Acta·Natasha S BartenevaIvan A Vorobjev
May 28, 2015·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·C Bruce BagwellFrederic I Preffer
Oct 24, 2012·Cytometry. Part B, Clinical Cytometry·Frederic I Preffer
Jun 22, 2013·Cytometry. Part B, Clinical Cytometry·R Clive Landis
Sep 12, 2013·Cytometry. Part B, Clinical Cytometry·Shabnam TangriUNKNOWN ICSH/ICCS Working Group
Jul 24, 2014·Biochemistry and Molecular Biology Education : a Bimonthly Publication of the International Union of Biochemistry and Molecular Biology·Laura E Ott, Susan Carson
Apr 9, 2015·Cytometry. Part B, Clinical Cytometry·C Bruce BagwellFrederic I Preffer

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