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Evaluating eosin-5-maleimide binding as a diagnostic test for hereditary spherocytosis in newborn infants

Journal of Perinatology volume 35, pages 357–361 (2015)Cite this article

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Abstract

Objective:

Neonates with undiagnosed hereditary spherocytosis (HS) are at risk for developing hazardous hyperbilirubinemia and anemia. Making an early diagnosis of HS in a neonate can prompt anticipatory guidance to prevent these adverse outcomes. A recent comparison study showed that a relatively new diagnostic test for HS, eosin-5-maleimide (EMA)-flow cytometry, performs better than other available tests in confirming HS. However, reports have not specifically examined the performance of this test among neonates.

Study design:

We compared EMA-flow cytometry from blood samples of healthy control neonates vs samples from neonates suspected of having HS on the basis of severe Coombs-negative jaundice and spherocytes on blood film. The diagnosis of HS was later either confirmed or excluded based on clinical findings and next generation sequencing (NGS) after which we correlated the EMA-flow results with the diagnosis.

Result:

EMA-flow was performed on the blood of 31 neonates; 20 healthy term newborns and 11 who were suspected of having HS. Eight of the 11 were later confirmed positive for HS and one was confirmed positive for hereditary elliptocytosis (HE). All nine had persistently abnormal erythroid morphology, reticulocytosis and anemia, and eight of the nine had relevant mutations discovered using NGS. The other was confirmed positive for HS on the basis that a parent had HS, and the neonate’s spherocytosis, reticulocytosis and anemia persisted. The 20 healthy controls and the 2 in whom HS was initially suspected but later excluded all had EMA-flow results in the range reported in healthy children and adults. In contrast, all nine in whom HS or HE was confirmed had abnormal EMA-flow results consistent with previous reports in older children and adults with HS.

Conclusion:

Although our sample size is small, our findings are consistent with the literature in older children and adults suggesting that EMA-flow cytometric testing performs well in supporting the diagnosis of HS/HE during the early neonatal period.

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References

  1. Johnson L, Bhutani VK, Karp K, Sivieri EM, Shapiro SM . Clinical report from the pilot USA kernicterus registry (1992–2004). J Perinatol 2009; 29 (Suppl 1): S25–S45.

    Article  Google Scholar 

  2. Christensen RD, Yaish HM, Nussenzveig RH, Reading NS, Agarwal AM, Eggert LD et al. Acute kernicterus in a neonate with O/B blood group incompatibility and a mutation in SLC4A1. Pediatrics 2013; 132 (2): e531–e534.

    Article  Google Scholar 

  3. Gallagher PG . Abnormalities of the erythrocyte membrane. Pediatr Clin North Am 2013; 60 (6): 1349–1362.

    Article  Google Scholar 

  4. Christensen RD, Henry E . Hereditary spherocytosis in neonates with hyperbilirubinemia. Pediatrics 2010; 125 (1): 120–125.

    Article  Google Scholar 

  5. Sheffield MJ, Christensen RD . Evaluating neonatal hyperbilirubinemia in late preterm Hispanic twins led to the diagnosis of hereditary spherocytosis in them, and in their sibling and in their mother. J Perinatol 2011; 31 (9): 625–627.

    Article  CAS  Google Scholar 

  6. Christensen RD, Yaish HM, Lemons RS . Neonatal hemolytic jaundice: Morphologic features of erythrocytes that will help you diagnose the underlying condition. Neonatology 2014; 105 (4): 243–249.

    Article  CAS  Google Scholar 

  7. Bianchi P, Fermo E, Vercellati C, Marcello AP, Porretti L, Cortelezzi A et al. Diagnostic power of laboratory tests for hereditary spherocytosis: a comparison study in 150 patients grouped according to molecular and clinical characteristics. Haematologica 2012; 97 (4): 516–523.

    Article  CAS  Google Scholar 

  8. Kedar PS, Colah RB, Kulkarni S, Ghosh K, Mohanty D . Experience with eosin-5'-maleimide as a diagnostic tool for red cell membrane cytoskeleton disorders. Clin Lab Haematol 2003; 25 (6): 373–376.

    Article  CAS  Google Scholar 

  9. King MJ, Smythe JS, Mushens R . Eosin-5-maleimide binding to band 3 and Rh-related proteins forms the basis of a screening test for hereditary spherocytosis. Br J Haematol 2004; 124: 106–113.

    Article  CAS  Google Scholar 

  10. King MJ, Telfer P, MacKinnon H, Langabeer L, McMahon C, Darbyshire P et al. Using the eosin-5-maleimide binding test in the differential diagnosis of hereditary spherocytosis and hereditary pyropoikilocytosis. Cytometry B Clin Cytom 2008; 74 (4): 244–250.

    Article  Google Scholar 

  11. Kar R, Mishra P, Pati HP . Evaluation of eosin-5-maleimide flow cytometric test in diagnosis of hereditary spherocytosis. Int J Lab Hematol 2010; 32 (1 Pt 2): 8–16.

    Article  CAS  Google Scholar 

  12. Kar R, Rao S, Srinivas UM, Mishra P, Pati HP . Clinico-hematological profile of hereditary spherocytosis: experience from a tertiary care center in North India. Hematology 2009; 14 (3): 164–167.

    Article  Google Scholar 

  13. Tachavanich K, Tanphaichitr VS, Utto W, Viprakasit V . Rapid flow cytometric test using eosin-5-maleimide for diagnosis of red blood cell membrane disorders. Southeast Asian J Trop Med Public Health 2009; 40 (3): 570–575.

    CAS  PubMed  Google Scholar 

  14. King MJ, Jepson MA, Guest A, Mushens R . Detection of hereditary pyropoikilocytosis by the eosin-5-maleimide (EMA)-binding test is attributable to a marked reduction in EMA-reactive transmembrane proteins. Int J Lab Hematol 2011; 33 (2): 205–211.

    Article  Google Scholar 

  15. D'Alcamo E, Agrigento V, Sclafani S, Vitrano A, Cuccia L, Maggio A et al. Reliability of EMA binding test in the diagnosis of hereditary spherocytosis in Italian patients. Acta Haematol 2011; 125 (3): 136–140.

    Article  Google Scholar 

  16. Ciepiela O, Kotuła I, Górska E, Stelmaszczyk-Emmel A, Popko K, Szmydki-Baran A et al. Delay in the measurement of eosin-5′-maleimide (EMA) binding does not affect the test result for the diagnosis of hereditary spherocytosis. Clin Chem Lab Med 2013; 51 (4): 817–823.

    Article  CAS  Google Scholar 

  17. Golafshan HA, Ranjbaran R, Kalantari T, Moezzi L, Karimi M, Behzad-Behbahani A et al. Evaluation of red cell membrane cytoskeletal disorders using a flow cytometric method in South Iran. Turk J Haematol 2014; 31 (1): 25–31.

    Article  Google Scholar 

  18. Watanabe T, Ono H, Tajima I, Ishigaki H, Hakamata A, Shirai M et al. Hereditary spherocytosis diagnosed with the eosin-5'-maleimide binding test. Pediatr Int 2014; 56 (3): 427–429.

    Article  Google Scholar 

  19. Christensen RD, Nussenzveig RH, Yaish HM, Henry E, Eggert LD, Agarwal AM . Causes of hemolysis in neonates with extreme hyperbilirubinemia. J Perinatol 2014; 34 (8): 616–619.

    Article  CAS  Google Scholar 

  20. Henry E, Christensen RD, Sheffield MJ, Eggert LD, Carroll PD, Minton SD et al. Why do four NICUs using identical RBC transfusion guidelines have different gestational age-adjusted RBC transfusion rates? J Perinatol (epub ahead of print 25 September 2014).

  21. Delhommeau F, Cynober T, Schischmanoff PO, Rohrlich P, Delaunay J, Mohandas N et al. Natural history of hereditary spherocytosis during the first year of life. Blood 2000; 95: 393–397.

    CAS  PubMed  Google Scholar 

  22. Gallagher PG . Disorders of red cell volume regulation. Curr Opin Hematol 2013; 20 (3): 201–207.

    Article  CAS  Google Scholar 

  23. Bogardus HH, Maksimova YD, Forget BG, Gallagher PG . A de novo band 3 mutation in hereditary spherocytosis. Pediatr Blood Cancer 2012; 58 (6): 1004.

    Article  Google Scholar 

  24. Sangerman J, Maksimova Y, Edelman EJ, Morrow JS, Forget BG, Gallagher PG . Ankyrin-linked hereditary spherocytosis in an African-American kindred. Am J Hematol 2008; 83 (10): 789–794.

    Article  CAS  Google Scholar 

  25. Gallagher PG . Hematologically important mutations: ankyrin variants in hereditary spherocytosis. Blood Cells Mol Dis 2005; 35 (3): 345–347.

    Article  CAS  Google Scholar 

  26. Gundel F, Eber S, Heep A . A new ankyrin mutation (ANK1 EXON E9X) causing severe hereditary spherocytosis in the neonatal period. Ann Hematol 2011; 90 (2): 231–232.

    Article  Google Scholar 

  27. Berardi A, Lugli L, Ferrari F, Gargano G, D'Apolito M, Marrone A et al. Kernicterus associated with hereditary spherocytosis and UGT1A1 promoter polymorphism. Biol Neonate 2006; 90 (4): 243–246.

    Article  Google Scholar 

  28. Saada V, Cynober T, Brossard Y, Schischmanoff PO, Sender A, Cohen H et al. Incidence of hereditary spherocytosis in a population of jaundiced neonates. Pediatr Hematol Oncol 2006; 23 (5): 387–397.

    Article  Google Scholar 

  29. Crisp RL, Solari L, Vota D, Garcia E, Miguez G, Chamorro ME et al. A prospective study to assess the predictive value for hereditary spherocytosis using five laboratory tests (cryohemolysis test, eosin-5’-maleimide flow cytometry, osmotic fragility tests, autohemolysis test, and SDS-PAGE) on 50 hereditary spherocytosis families in Argentina. Ann Hematol 2011; 90 (6): 625–634.

    Article  Google Scholar 

  30. Girodon F, Garcon L, Bergoin E, Larger M, Delaunay J, Feneant-Thibault M et al. Usefulness of the eosin-5’maleimide cytometric method as a first-line screening test for the diagnosis of hereditary spherocytosis: comparison with ektacytometry and protein electrophoresis. Br J Haematol 2008; 140 (4): 468–470.

    Article  CAS  Google Scholar 

  31. Yaish HM, Christensen RD, Henry E, Baer VL, Bennett S . A simple method of screening newborn infants for hereditary spherocytosis. J Applied Hematol 2013; 4: 27–32.

    Google Scholar 

  32. Yaish HM, Christensen RD, Agarwal A . A neonate with Coombs-negative hemolytic jaundice with spherocytes but normal erythrocyte indices: a rare case of autosomal-recessive hereditary spherocytosis due to alpha-spectrin deficiency. J Perinatol 2013; 33 (5): 404–406.

    Article  CAS  Google Scholar 

  33. Nussenzveig RH, Christensen RD, Prchal JT, Yaish HM, Agrawal AM . Novel alpha spectrin mutation in trans with alpha spectrinLEPRA resulting in severe, non-dominant, hereditary spherocytosis. Neonatology 2014; 105 (4): 355–357.

    Article  Google Scholar 

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Author information

Authors and Affiliations

  1. The Department of Women and Newborns, Intermountain Healthcare, Salt Lake City, UT, USA,

    R D Christensen

  2. Division of Neonatology, University of Utah School of Medicine, Salt Lake City, UT, USA,

    R D Christensen

  3. Division of Hematology/Oncology, Department of Pediatrics, University of Utah School of Medicine, Salt Lake City, UT, USA,

    R D Christensen & H M Yaish

  4. Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA,

    A M Agarwal, R H Nussenzveig, N Heikal & M A Liew

  5. ARUP Laboratories, Salt Lake City, UT, USA

    A M Agarwal & N Heikal

Authors
  1. R D Christensen
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  4. N Heikal
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  6. H M Yaish
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Corresponding author

Correspondence to R D Christensen.

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Competing interests

Drs. Agarwal, Nussenzveig, Heikal and Liew are employees of ARUP Laboratories, the reference laboratory where the EMA-flow testing is performed. However, none are equity holders in the company and none benefit financially from the test. The other authors (RDC and HMY) have no conflicts of interest to disclose.

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Christensen, R., Agarwal, A., Nussenzveig, R. et al. Evaluating eosin-5-maleimide binding as a diagnostic test for hereditary spherocytosis in newborn infants. J Perinatol 35, 357–361 (2015). https://doi.org/10.1038/jp.2014.202

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