Blood Res 2018; 53(1): 10-17
Cryohemolysis, erythrocyte osmotic fragility, and supplementary hematimetric indices in the diagnosis of hereditary spherocytosis
Ledesma Achem Miryam Emilse1, Haro Cecilia1, Terán Magdalena María1, Mónaco María Eugenia2, Issé Blanca Alicia1, Sandra Stella Lazarte1

1Instituto de Bioquímica Aplicada, Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Tucumán, Argentina.

2Instituto de Biología, Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Tucumán, Argentina.

Correspondence to: Sandra Stella Lazarte, Ph.D. Instituto de Bioqu?mica Aplicada, Universidad Nacional de Tucum?n, Balcarce 747, San Miguel de Tucum?, Tucum?n, CP4000 Argentina.
Received: June 13, 2017; Revised: July 20, 2017; Accepted: August 30, 2017; Published online: March 31, 2018.
© The Korean Journal of Hematology. All rights reserved.

cc This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.


Hereditary spherocytosis (HS) is a chronic hemolytic anemia characterized by microspherocytes in the peripheral blood and increased erythrocyte osmotic fragility (EOF). This study evaluated the cryohemolysis test (CHT); initial hemolysis (IH); immediate and incubated hemolysis percentage in 5.5 g/L NaCl (H5.5); mean corpuscular hemoglobin concentration (MCHC); red blood cell distribution width (RDW); and Hb/MCHC, Hb/RDW, and MCHC/RDW ratios for the diagnosis of HS.


Data from 13 patients with HS were evaluated at the Instituto de Bioqu?mica Aplicada and compared with data from 14 unaffected individuals and 11 patients with anemia due to another etiology. Total blood and reticulocyte counts, CHT, and immediate and incubated EOF were performed in all subjects; sensitivity, specificity, efficiency, and Youden index (YI) were calculated.


Eight patients with HS had MCHC ≥345 g/L, 10 had RDW ≥14.5%, 12 had IH >5.0 g/L, 11 had immediate H5.5 ≥5%, and 13 had incubated H5.5 ≥50% (the cut-off value to consider HS). The efficiency and YI were: immediate H5.5 (0.94?0.85), incubated H5.5 (0.89?0.82), IH (0.89?0.78), MCHC (0.87?0.62), CHT (0.84?0.54), and Hb/MCHC (0.71?0.56), respectively. The calculated ratios could distinguish subjects with HS from unaffected individuals (P<0.05), but not those with anemia of another etiology (P>0.05).


Although the CHT and supplementary hematimetric indexes were useful to differentiate individuals with SH from healthy controls, they cannot distinguish from anemias of other etiology. CHT and MCHC, in addition to EOF, are recommended for diagnosing HS patients because of their low cost and efficiency.

Keywords: Hereditary spherocytosis, Cryohemolysis, Erythrocyte osmotic fragility, Hematimetric indexes


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