Flow cytometric osmotic fragility test and eosin‐5’‐maleimide dye‐binding tests are better than conventional osmotic fragility tests for the diagnosis of hereditary spherocytosis

Introduction Hereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico‐laboratory manifestations. We evaluated the flow‐cytometric tests: eosin‐5’‐maleimide (EMA) and flow‐cytometric osmotic fragility test (FOFT) and the conventional osmotic fragility tes...

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Published in:	International journal of laboratory hematology Vol. 40; no. 3; pp. 335 - 342
Main Authors:	Arora, R. D., Dass, J., Maydeo, S., Arya, V., Radhakrishnan, N., Sachdeva, A., Kotwal, J., Bhargava, M.
Format:	Journal Article
Language:	English
Published:	England Wiley Subscription Services, Inc 01.06.2018
Subjects:	Adult Aged Anemia Diagnosis Dyes Eosine Yellowish-(YS) - analogs & derivatives Eosine Yellowish-(YS) - metabolism eosin‐5’‐maleimide eosin‐5’‐maleimide decrease Erythrocytes flow cytometric Flow cytometry Flow Cytometry - methods flow osmotic fragility tests Hereditary spherocytosis Humans Immune hemolytic anemia Middle Aged Osmotic Fragility residual red cells Sensitivity and Specificity Spherocytosis Spherocytosis, Hereditary - diagnosis eosin-5’-maleimide decrease eosin-5’-maleimide flow cytometric residual red cells flow osmotic fragility tests Hereditary spherocytosis
ISSN:	1751-5521, 1751-553X, 1751-553X
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Abstract	Introduction Hereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico‐laboratory manifestations. We evaluated the flow‐cytometric tests: eosin‐5’‐maleimide (EMA) and flow‐cytometric osmotic fragility test (FOFT) and the conventional osmotic fragility tests (OFT) for the diagnosis of hereditary spherocytosis (HS). Methods One hundred two suspected HS patients underwent EMA, FOFT, incubated OFT (IOFT), and room temperature OFT (RT‐OFT). In addition, 10 cases of immune hemolytic anemia (IHA) were included, and performance of the above 4 tests was evaluated. For EMA and FOFT, 5 normal controls were assessed together with the patients and cutoffs were calculated using receiver‐operator‐characteristics curve (ROC) analysis. Results The best cutoff for %EMA decrease was 12.5%, and for FOFT, %residual red cells (%RRC) was 25.6%. The sensitivity and specificity of RT‐OFT was 62.06% and 86.3%, respectively, while that of IOFT was 79.31% and 87.67%, respectively. Both flow cytometric tests performed better. Sensitivity and specificity of EMA was 86.2% and 93.9% respectively, and that of FOFT was 96.6% and 98.63%, respectively. The combination of the FOFT with IOFT or EMA dye‐binding test yields a sensitivity of 100%, but with EMA, it had a higher specificity. Hb/MCHC was a predictor of the severity of the disease while %EMA decrease and %RRC did not correlate with severity of the disease. Conclusion Flow‐cytometric osmotic fragility test is the best possible single test followed by EMA for diagnosis of HS. A combination of FOFT and EMA can correctly diagnose 100% patients. These tests are likely to replace conventional OFTs in future.
AbstractList	Hereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico-laboratory manifestations. We evaluated the flow-cytometric tests: eosin-5'-maleimide (EMA) and flow-cytometric osmotic fragility test (FOFT) and the conventional osmotic fragility tests (OFT) for the diagnosis of hereditary spherocytosis (HS).INTRODUCTIONHereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico-laboratory manifestations. We evaluated the flow-cytometric tests: eosin-5'-maleimide (EMA) and flow-cytometric osmotic fragility test (FOFT) and the conventional osmotic fragility tests (OFT) for the diagnosis of hereditary spherocytosis (HS).One hundred two suspected HS patients underwent EMA, FOFT, incubated OFT (IOFT), and room temperature OFT (RT-OFT). In addition, 10 cases of immune hemolytic anemia (IHA) were included, and performance of the above 4 tests was evaluated. For EMA and FOFT, 5 normal controls were assessed together with the patients and cutoffs were calculated using receiver-operator-characteristics curve (ROC) analysis.METHODSOne hundred two suspected HS patients underwent EMA, FOFT, incubated OFT (IOFT), and room temperature OFT (RT-OFT). In addition, 10 cases of immune hemolytic anemia (IHA) were included, and performance of the above 4 tests was evaluated. For EMA and FOFT, 5 normal controls were assessed together with the patients and cutoffs were calculated using receiver-operator-characteristics curve (ROC) analysis.The best cutoff for %EMA decrease was 12.5%, and for FOFT, %residual red cells (%RRC) was 25.6%. The sensitivity and specificity of RT-OFT was 62.06% and 86.3%, respectively, while that of IOFT was 79.31% and 87.67%, respectively. Both flow cytometric tests performed better. Sensitivity and specificity of EMA was 86.2% and 93.9% respectively, and that of FOFT was 96.6% and 98.63%, respectively. The combination of the FOFT with IOFT or EMA dye-binding test yields a sensitivity of 100%, but with EMA, it had a higher specificity. Hb/MCHC was a predictor of the severity of the disease while %EMA decrease and %RRC did not correlate with severity of the disease.RESULTSThe best cutoff for %EMA decrease was 12.5%, and for FOFT, %residual red cells (%RRC) was 25.6%. The sensitivity and specificity of RT-OFT was 62.06% and 86.3%, respectively, while that of IOFT was 79.31% and 87.67%, respectively. Both flow cytometric tests performed better. Sensitivity and specificity of EMA was 86.2% and 93.9% respectively, and that of FOFT was 96.6% and 98.63%, respectively. The combination of the FOFT with IOFT or EMA dye-binding test yields a sensitivity of 100%, but with EMA, it had a higher specificity. Hb/MCHC was a predictor of the severity of the disease while %EMA decrease and %RRC did not correlate with severity of the disease.Flow-cytometric osmotic fragility test is the best possible single test followed by EMA for diagnosis of HS. A combination of FOFT and EMA can correctly diagnose 100% patients. These tests are likely to replace conventional OFTs in future.CONCLUSIONFlow-cytometric osmotic fragility test is the best possible single test followed by EMA for diagnosis of HS. A combination of FOFT and EMA can correctly diagnose 100% patients. These tests are likely to replace conventional OFTs in future. Hereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico-laboratory manifestations. We evaluated the flow-cytometric tests: eosin-5'-maleimide (EMA) and flow-cytometric osmotic fragility test (FOFT) and the conventional osmotic fragility tests (OFT) for the diagnosis of hereditary spherocytosis (HS). One hundred two suspected HS patients underwent EMA, FOFT, incubated OFT (IOFT), and room temperature OFT (RT-OFT). In addition, 10 cases of immune hemolytic anemia (IHA) were included, and performance of the above 4 tests was evaluated. For EMA and FOFT, 5 normal controls were assessed together with the patients and cutoffs were calculated using receiver-operator-characteristics curve (ROC) analysis. The best cutoff for %EMA decrease was 12.5%, and for FOFT, %residual red cells (%RRC) was 25.6%. The sensitivity and specificity of RT-OFT was 62.06% and 86.3%, respectively, while that of IOFT was 79.31% and 87.67%, respectively. Both flow cytometric tests performed better. Sensitivity and specificity of EMA was 86.2% and 93.9% respectively, and that of FOFT was 96.6% and 98.63%, respectively. The combination of the FOFT with IOFT or EMA dye-binding test yields a sensitivity of 100%, but with EMA, it had a higher specificity. Hb/MCHC was a predictor of the severity of the disease while %EMA decrease and %RRC did not correlate with severity of the disease. Flow-cytometric osmotic fragility test is the best possible single test followed by EMA for diagnosis of HS. A combination of FOFT and EMA can correctly diagnose 100% patients. These tests are likely to replace conventional OFTs in future. Introduction Hereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico‐laboratory manifestations. We evaluated the flow‐cytometric tests: eosin‐5’‐maleimide (EMA) and flow‐cytometric osmotic fragility test (FOFT) and the conventional osmotic fragility tests (OFT) for the diagnosis of hereditary spherocytosis (HS). Methods One hundred two suspected HS patients underwent EMA, FOFT, incubated OFT (IOFT), and room temperature OFT (RT‐OFT). In addition, 10 cases of immune hemolytic anemia (IHA) were included, and performance of the above 4 tests was evaluated. For EMA and FOFT, 5 normal controls were assessed together with the patients and cutoffs were calculated using receiver‐operator‐characteristics curve (ROC) analysis. Results The best cutoff for %EMA decrease was 12.5%, and for FOFT, %residual red cells (%RRC) was 25.6%. The sensitivity and specificity of RT‐OFT was 62.06% and 86.3%, respectively, while that of IOFT was 79.31% and 87.67%, respectively. Both flow cytometric tests performed better. Sensitivity and specificity of EMA was 86.2% and 93.9% respectively, and that of FOFT was 96.6% and 98.63%, respectively. The combination of the FOFT with IOFT or EMA dye‐binding test yields a sensitivity of 100%, but with EMA, it had a higher specificity. Hb/MCHC was a predictor of the severity of the disease while %EMA decrease and %RRC did not correlate with severity of the disease. Conclusion Flow‐cytometric osmotic fragility test is the best possible single test followed by EMA for diagnosis of HS. A combination of FOFT and EMA can correctly diagnose 100% patients. These tests are likely to replace conventional OFTs in future. IntroductionHereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico‐laboratory manifestations. We evaluated the flow‐cytometric tests: eosin‐5’‐maleimide (EMA) and flow‐cytometric osmotic fragility test (FOFT) and the conventional osmotic fragility tests (OFT) for the diagnosis of hereditary spherocytosis (HS).MethodsOne hundred two suspected HS patients underwent EMA, FOFT, incubated OFT (IOFT), and room temperature OFT (RT‐OFT). In addition, 10 cases of immune hemolytic anemia (IHA) were included, and performance of the above 4 tests was evaluated. For EMA and FOFT, 5 normal controls were assessed together with the patients and cutoffs were calculated using receiver‐operator‐characteristics curve (ROC) analysis.ResultsThe best cutoff for %EMA decrease was 12.5%, and for FOFT, %residual red cells (%RRC) was 25.6%. The sensitivity and specificity of RT‐OFT was 62.06% and 86.3%, respectively, while that of IOFT was 79.31% and 87.67%, respectively. Both flow cytometric tests performed better. Sensitivity and specificity of EMA was 86.2% and 93.9% respectively, and that of FOFT was 96.6% and 98.63%, respectively. The combination of the FOFT with IOFT or EMA dye‐binding test yields a sensitivity of 100%, but with EMA, it had a higher specificity. Hb/MCHC was a predictor of the severity of the disease while %EMA decrease and %RRC did not correlate with severity of the disease.ConclusionFlow‐cytometric osmotic fragility test is the best possible single test followed by EMA for diagnosis of HS. A combination of FOFT and EMA can correctly diagnose 100% patients. These tests are likely to replace conventional OFTs in future.
Author	Kotwal, J. Arora, R. D. Maydeo, S. Sachdeva, A. Dass, J. Arya, V. Radhakrishnan, N. Bhargava, M.
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Keywords	eosin-5’-maleimide decrease eosin-5’-maleimide flow cytometric residual red cells flow osmotic fragility tests Hereditary spherocytosis
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Snippet	Introduction Hereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico‐laboratory manifestations. We evaluated the... Hereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico-laboratory manifestations. We evaluated the... IntroductionHereditary spherocytosis (HS) is the most common inherited hemolytic anemia with heterogeneous clinico‐laboratory manifestations. We evaluated the...
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SubjectTerms	Adult Aged Anemia Diagnosis Dyes Eosine Yellowish-(YS) - analogs & derivatives Eosine Yellowish-(YS) - metabolism eosin‐5’‐maleimide eosin‐5’‐maleimide decrease Erythrocytes flow cytometric Flow cytometry Flow Cytometry - methods flow osmotic fragility tests Hereditary spherocytosis Humans Immune hemolytic anemia Middle Aged Osmotic Fragility residual red cells Sensitivity and Specificity Spherocytosis Spherocytosis, Hereditary - diagnosis
Title	Flow cytometric osmotic fragility test and eosin‐5’‐maleimide dye‐binding tests are better than conventional osmotic fragility tests for the diagnosis of hereditary spherocytosis
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