Importância e vantagem da citometria de fluxo frente aos testes de triagem no diagnóstico da hemoglobinúria paroxística noturna

Modesto, Thaís M.; Neves, Maria Amélia; Brito, Ana Elita de; Araújo, Rosane C. P.; Santos, Nilcilene Nascimento dos; Valgueiro, Maria do Carmo; Machado, Cíntia

doi:10.1590/s1516-84842006000400011

Cited by 4 publications

(3 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Blood transfusions performed in the months preceding material collection were not a restriction to patient participation, as they do not interfere in the analysis of leukocyte populations (11) . The control group was formed with 20 adult volunteers (13 women, 7 men, mean age: 44 years), with no hematological diseases.…”

Section: Methodsmentioning

confidence: 99%

Detection of PNH cells by flow cytometry, using multiparameter analysis

Rocha¹,

Silva²,

Souza³

et al. 2014

Jornal Brasileiro De Patologia E Medicina Laboratorial

View full text Add to dashboard Cite

Introduction: The laboratory diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), disease that is categorized by reduced synthesis of glycosylphosphatidylinositol (GPI) anchor, is based on the detection of blood cells deficient in GPI-anchored proteins by flow cytometry. PNH clones have been detected in patients with aplastic anaemia (AA) and myelodysplastic syndrome (MDS), with therapeutic implications. Objectives: To validate a sensitive assay for detection of GPI-anchored protein-deficient cells, by flow cytometry, and to analyze the clone frequency in AA and MDS patients. Methods: Samples from 20 AA patients, 30 MDS patients and 20 adult volunteers (control group) were analyzed using monoclonal antibodies to CD16, CD24, CD55 and CD59 (neutrophils); CD14 and CD55 (monocytes); CD55 and CD59 (erythrocytes); besides fluorescent aerolysin reagent (FLAER) (neutrophils and monocytes) and lineage markers. The proportions of PNH cells detected in neutrophils and monocytes, using different reagent combinations, were compared by analysis of variance (ANOVA) and Pearson's correlation. Results: PNH cells were detected in five (25%) AA patients, and the proportions of PNH cells varied from 0.14% to 94.84% of the analyzed events. PNH cells were not detected in the MDS patients. However, by the analysis of these samples, it was possible to identify the technical challenges caused by the presence of immature and dysplastic circulating cells. FLAER showed clear distinction of GPI-deficient cells. Conclusion: Multiparameter flow cytometry analysis offers high sensitivity and accuracy in the detection of subclinical PNH clones. FLAER shows excellent performance in detection of PNH neutrophils and monocytes.

show abstract

Section: Methodsmentioning

confidence: 99%

Detection of PNH cells by flow cytometry, using multiparameter analysis

Rocha¹,

Silva²,

Souza³

et al. 2014

Jornal Brasileiro De Patologia E Medicina Laboratorial

View full text Add to dashboard Cite

show abstract

“…Rarely, spontaneous remission occurs (10% to 15% of cases) or transformation to acute myeloid leukemia. (3,9,13,14) …”

Section: Introductionmentioning

confidence: 99%

Frequency of paroxysmal nocturnal hemoglobinuria in patients attended in Belém, Pará, Brazil

Junior¹,

Cardoso²,

Rocha³

et al. 2010

Rev. Bras. Hematol. Hemoter.

View full text Add to dashboard Cite

Background Paroxysmal nocturnal hemoglobinuria is a hematological disease with complex physiopathology. It is genetically characterized by a somatic mutation in the PIG-A gene (phosphatidylinositol glycan anchor biosynthesis, class A), in which the best known antigens are DAF (decay accelerating factor or CD55) and MIRL (membrane inhibitor of reactive lysis or CD59). Objective To determine the frequency of paroxysmal nocturnal hemoglobinuria in patients attended at the HEMOPA foundation from November 2008 to July 2009. Method Thirty patients, with ages ranging from two to 79 years old and suspected of having paroxysmal nocturnal hemoglobinuria were examined. All patients were immunophenotyped by flow cytometry for the CD5, CD59, CD16 and CD45 antigens. Results Paroxysmal nocturnal hemoglobinuria was identified in nine of the thirty patients investigated. Another 3 cases had inconclusive results with CD59-negative labeling only for neutrophils. The highest frequency of paroxysmal nocturnal hemoglobinuria patients (7/9) and inconclusive cases (2/3) were between 19 years old and 48 years old, with a median of 28 years. Conclusion These results show the importance of flow cytometry to identify cases in which patients are deficient in only one antigen (CD59).

show abstract

“…Rarely, spontaneous remission occurs (10% to 15% of cases) or transformation to acute myeloid leukemia. (3,9,13,14) The diagnosis of PNH is based on clinical findings and laboratory tests that detect GPI-binding to red cell membrane proteins or that demonstrate the presence of red blood cells that are exceptionally sensitive to hemolysis by the complement system. These tests include the acidified serum test (Ham's test), determination of the CD55 and CD59 antigens using gel column technology or immunophenotyping by flow cytometry.…”

Section: Introductionmentioning

confidence: 99%