Comparison of High Sensitivity and Conventional Flow Cytometry for Diagnosing Overt Paroxysmal Nocturnal Hemoglobinuria and Detecting Minor Paroxysmal Nocturnal Hemoglobinuria Clones
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Background: High sensitivity flow cytometry (HS-FCM) was recently developed for diagnosing paroxysmal nocturnal hemoglobinuria (PNH). We compared its performance with conventional flow cytometry (C-FCM) for diagnosing overt PNH and detecting minor (0.1–1%) PNH clones in aplastic anemia (AA)/low-grade myelodysplastic syndrome (MDS) patients. Methods: C-FCM and HS-FCM were performed simultaneously on 41 samples from healthy controls and 23 peripheral blood samples from 15 AA/low-grade MDS and eight PNH patients, using a Navios flow cytometer (Beckman Coulter, Miami, FL, USA). Results were compared. Results: No healthy control samples had PNH clone size >0.01%. For granulocytes, C-FCM detected a smaller PNH clone size than HS-FCM (mean difference: 0.7–1.7%). In AA/low-grade MDS patients, three samples showed >1% PNH clones with C-FCM but not with HS-FCM. Seven samples showed minor PNH clones by C-FCM, but HS-FCM showed negative results for all these samples. In PNH patients, C-FCM detected a smaller PNH clone size than HS-FCM (mean difference: 1.9–5.0%). For red blood cells, C-FCM detected a greater PNH clone size than HS-FCM (mean difference: 1.5%). In AA/low-grade MDS patients, C-FCM showed >1% PNH clones in six samples, but HS-FCM showed >1% PNH clones in none of the samples. C-FCM detected minor PNH clones in nine samples, but six of them were negative by HS-FCM. In PNH patients, C-FCM detected a greater PNH clone size than HS-FCM (mean difference: 2.5%). Conclusions: HS-FCM can sensitively detect minor PNH clones and reduce false-positive C-FCM minor PNH clone cases in AA/low-grade MDS patients.
Abstract
Background: High sensitivity flow cytometry (HS-FCM) was recently developed for diagnosing paroxysmal nocturnal hemoglobinuria (PNH). We compared its performance with conventional flow cytometry (C-FCM) for diagnosing overt PNH and detecting minor (0.1–1%) PNH clones in aplastic anemia (AA)/low-grade myelodysplastic syndrome (MDS) patients. Methods: C-FCM and HS-FCM were performed simultaneously on 41 samples from healthy controls and 23 peripheral blood samples from 15 AA/low-grade MDS and eight PNH patients, using a Navios flow cytometer (Beckman Coulter, Miami, FL, USA). Results were compared. Results: No healthy control samples had PNH clone size >0.01%. For granulocytes, C-FCM detected a smaller PNH clone size than HS-FCM (mean difference: 0.7–1.7%). In AA/low-grade MDS patients, three samples showed >1% PNH clones with C-FCM but not with HS-FCM. Seven samples showed minor PNH clones by C-FCM, but HS-FCM showed negative results for all these samples. In PNH patients, C-FCM detected a smaller PNH clone size than HS-FCM (mean difference: 1.9–5.0%). For red blood cells, C-FCM detected a greater PNH clone size than HS-FCM (mean difference: 1.5%). In AA/low-grade MDS patients, C-FCM showed >1% PNH clones in six samples, but HS-FCM showed >1% PNH clones in none of the samples. C-FCM detected minor PNH clones in nine samples, but six of them were negative by HS-FCM. In PNH patients, C-FCM detected a greater PNH clone size than HS-FCM (mean difference: 2.5%). Conclusions: HS-FCM can sensitively detect minor PNH clones and reduce false-positive C-FCM minor PNH clone cases in AA/low-grade MDS patients.
참고문헌 (18)
- Miyata T, "The cloning of PIG-A, a component in the early step of GPI-anchor biosynthesis", Science, 259, pp.1318~1320 (1993)
- Takeda J, "Deficiency of the GPI anchor caused by a somatic mutation of the PIG-A gene in paroxysmal nocturnal hemoglobinuria", Cell, 73, pp.703~711 (1993)
- Hillmen P, "Natural history of paroxysmal nocturnal hemoglobinuria", N Engl J Med, 333, pp.1253~1258 (1995)
- Parker CJ, "Bone marrow failure syndromes : paroxysmal nocturnal hemoglobinuria", Hematol Oncol Clin North Am, 23, pp.333~346 (2009)
- Richards SJ, "Application of flow cytometry to the diagnosis of paroxysmal nocturnal hemoglobinuria", Cytometry, 42, pp.223~233 (2000)
- Raza A, "A prospective multicenter study of paroxysmal nocturnal hemoglobinuria cells in patients with bone marrow failure", Cytometry B Clin Cytom, 86, pp.175~182 (2014)
- Sugimori C, "Minor population of CD55–CD59- blood cells predicts response to immunosuppressive therapy and prognosis in patients with aplastic anemia", Blood, 107, pp.1308~1314 (2006)
- Dunn DE, "Paroxysmal nocturnal hemoglobinuria cells in patients with bone marrow failure syndromes", Ann Intern Med, 131, pp.401~408 (1999)
- Tichelli A, "Late clonal complications in severe aplastic anemia", Leuk Lymphoma, 12, pp.167~175 (1994)
- Parker C, "Diagnosis and management of paroxysmal nocturnal hemoglobinuria", Blood, 106, pp.3699~3709 (2005)
- Borowitz MJ, "Guidelines for the diagnosis and monitoring of paroxysmal nocturnal hemoglobinuria and related disorders by flow cytometry", Cytometry B Clin Cytom, 78, pp.211~230 (2010)
- Sutherland DR, "Practical guidelines for the high-sensitivity detection and monitoring of paroxysmal nocturnal hemoglobinuria clones by flow cytometry", Cytometry B Clin Cytom, 82, pp.195~208 (2012)
- Sutherland DR, "Use of a FLAER-based white blood cell assay in the primary screening of PNH clones", Am J Clin Pathol, 132, pp.564~572 (2009)
- Marinov I, "Intra-and interlaboratory variability of paroxysmal nocturnal hemoglobinuria testing by flow cytometry following the 2012 Practical Guidelines for high-sensitivity paroxysmal nocturnal hemoglobinuria testing", Cytometry B Clin Cytom, 84, pp.229~236 (2013)
- Sipol AA, "An inter-laboratory comparison of PNH clone detection by high-sensitivity flow cytometry in a Russian cohort", Hematology, 20, pp.31~38 (2015)
- Arber DA, "The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia", Blood, 127, pp.2391~2405 (2016)
- Illingworth A, "ICCS", ESCCA consensus guidelines to detect GPI-deficient cells in paroxysmal nocturnal hemoglobinuria(PNH)and related disorders part 3-data analysis, reporting and case studies, 2018, pp.94, 49~66
- Sutherland DR, "ICCS", ESCCA consensus guidelines to detect GPI-deficient cells in paroxysmal nocturnal hemoglobinuria(PNH)and related disorders part 2-reagent selection and assay optimization for high-sensitivity testing, 2018, pp.94, 23~48