Risk of hereditary spherocytosis misdiagnosis due to limited effective diagnostic methods
Keywords:
anemia, flow cytometry, fluorescence, hereditary, spherocytosisAbstract
Introduction: Hereditary spherocytosis is a prevalent congenital hemolytic erythrocyte membranopathy. Laboratory diagnosis is traditionally based on erythrocyte morphology, yet 20% of cases may lack visible spherocytes, leading to misdiagnosis. The Eosin-5'-maleimide binding assay has emerged as a reliable diagnostic method.
Objective: Evaluate the effectiveness of Eosin-5'-maleimide binding assay in diagnosing hereditary spherocytosis in pediatrics with negative-direct-antiglobulin-test-hemolytic anemia.
Methods: A cross-sectional study was conducted with 66 negative-direct-antiglobulin-test-hemolytic-anemic patients and 44 healthy controls. The percentage of spherocytes was scored from peripheral blood smear observation; the percentage of decreased mean fluorescent intensity from Eosin-5'-maleimide binding assay was used to determine red cell membrane protein deficiency, and hereditary spherocytosis was finally confirmed by gene sequencing as a gold standard.
Results: Eosin-5'-maleimide binding assay demonstrated high sensitivity (93.33%), specificity (100%), and accuracy (97.3%) in detecting hereditary spherocytosis, with an optimal cut-off value of 47.83% decreased mean fluorescent intensity.
Conclusion: The Eosin-5'-maleimide binding assay is a highly accurate diagnostic tool that outperforms traditional blood smear analysis. It is precious for detecting hereditary spherocytosis in patients with mild clinical manifestations or coexisting hemolytic disorders. This assay should be considered a frontline test for diagnosing hereditary spherocytosis in patients with hemolytic anemia.
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References
Wu C, Xiong T, Xu Z, Zhan C, Chen F, Ye Y, et al. Preliminary Study on the Clinical and Genetic Characteristics of Hereditary Spherocytosis in 15 Chinese Children [Internet]. Frontiers in Genetics. 2021;12. DOI: 10.3389/fgene.2021.652376
Asten I, Wit H, Oirschot B, Wijk R, Vries T. Hereditary spherocytosis without pronounced spherocytes on the peripheral blood smear [Internet]. International Journal of Laboratory Hematology. 2023; 45(1):6-8. DOI: 10.1111/ijlh.13976
Bello A. Hereditary Spherocytosis Misdiagnosed as Glucose-6-Phosphate Dehydrogenase Deficiency [Internet]. In: Tohid H, Baratta LG, Maibach H, editors. The Misdiagnosis Casebook in Clinical Medicine: A Case-Based Guide. Cham: Springer International Publishing; 2023. p. 275-82. DOI: 10.1007/978-3-031-28296-6_39
Sridevi HB. Combined Hereditary Spherocytosis and β-thalassemia trait: A Rare Co-existence [Internet]. Journal of Clinical and Diagnostic Research. 2018; 12(1):9-11. DOI: 10.7860/JCDR/2018/31302.11072
Alvarez Flores MB, Quintero V, Bravo A, Sevilla J, Huerta J, Pacheco S, et al. Diagnosis of hereditary spherocytosis by flow cytometry in paediatric patients. Multicentric study [Internet]. Clinica Chimica Acta. 2019; 493:S597. DOI: 10.1016/j.cca.2019.03.1252
Chueh HW, Hwang SM, Shim YJ, Lee JM, Park HS, Lee JH, et al. Korean clinical practice guidelines for the diagnosis of hereditary hemolytic anemia [Internet]. Blood Res. 2022; 57(2):86-94. DOI: 10.5045/br.2022.2021224
Wu Y, Liao L, Lin F. The diagnostic protocol for hereditary spherocytosis-2021 update [Internet]. JClin LabAnal. 2021; 35(12):e24034. DOI: 10.1002/jcla.24034
Tran DC, Dang AL, Hoang TNL, Nguyen CT, Le TMP, Dinh TNM, et al. Prevalence of Thalassemia in the Vietnamese Population and Building a Clinical Decision Support System for Prenatal Screening for Thalassemia [Internet]. Mediterr J Hematol Infect Dis. 2023; 15(1):e2023026. DOI: 10.4084/mjhid.2023.026
Palmer L, Briggs C, McFadden S, Zini G, Burthem J, Rozenberg G, et al. ICSH recommendations for the standardization of nomenclature and grading of peripheral blood cell morphological features [Internet]. Int J Lab Hematol. 2015; 37(3):287-303. DOI: 10.1111/ijlh.12327
Chase ML, Drews R, Zumberg MS, Ellis LR, Reid EG, Gerds AT, et al. Consensus recommendations on peripheral blood smear review: defining curricular standards and fellow competency [Internet]. Blood Advances. 2023; 7(13):3244-52. DOI: 10.1182/bloodadvances.2023009843
Arora RD, Dass J, Maydeo S, Arya V, Radhakrishnan N, Sachdeva A, et al. 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 [Internet]. Int J Lab Hematol. 2018; 40(3):335-42. DOI: 10.1111/ijlh.12794
Liu Y, Zheng J, Song L, Fang Y, Sun C, Li N, et al. A novel SPTB gene mutation in neonatal hereditary spherocytosis: A case report [Internet]. Exp Ther Med. 2020; 20(4):3253-9. DOI: 10.3892/etm.2020.9062
Choi HS, Choi Q, Kim JA, Im KO, Park SN, Park Y, et al. Molecular diagnosis of hereditary spherocytosis by multi-gene target sequencing in Korea: matching with osmotic fragility test and presence of spherocyte [Internet]. Orphanet J Rare Dis. 2019; 14(1):114. DOI: 10.1186/s13023-019-1070-0
King MJ, Garçon L, Hoyer JD, Iolascon A, Picard V, Stewart G, et al. ICSH guidelines for the laboratory diagnosis of nonimmune hereditary red cell membrane disorders [Internet]. International journal of laboratory hematology. 2015; 37(3):304-25. DOI: 10.1111/ijlh.12335
Yamamoto KS, Utshigisawa T, Ogura H, Aoki T, Kawakami T, Ohga S, et al. Clinical and genetic diagnosis of thirteen Japanese patients with hereditary spherocytosis [Internet]. Human Genome Variation. 2022; 9(1):1. DOI: 10.1038/s41439-021-00179-1
Kutter D. Hereditary spherocytosis is more frequent than expected: what to tell the patient? [Internet]. Bulletin de la Societe des Sciences Medicales du Grand-duche de Luxembourg. 2005; 12(1):7-22. DOI: 10.1002/jcla.24034
Christensen RD, Agarwal AM, Nussenzveig RH, Heikal N, Liew MA, Yaish HM. Evaluating eosin-5-maleimide binding as a diagnostic test for hereditary spherocytosis in newborn infants [Internet]. J Perinatol. 2015; 35(5):357-61. DOI: 10.1002/jcla.24034
Glenthøj A, Brieghel C, Nardo-Marino A, van Wijk R, Birgens H, Petersen J. Facilitating EMA binding test performance using fluorescent beads combined with next-generation sequencing [Internet]. EJHaem. 2021; 2(4):716-28. DOI: 10.1002/jha2.277
Park SH, Park C-J, Lee B-R, Kim YJ, Cho Y-U, Jang S, et al. Screening for hereditary spherocytosis: EMA binding test and flow cytometric osmotic fragility test are recommended, but cryohemolysis test is not recommended [Internet]. Blood. 2013; 122(21):3425. DOI: 10.1182/blood.V122.21.3425.3425
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