Hemolysis area: A new parameter of erythrocyte osmotic fragility for screening of thalassemia trait

Abstract

BACKGROUND: One-tube osmotic fragility test (OFT) is widely used for screening thalassemia traits. Interobserver variation may occur with 0.36% NaCl-based OFT due to the naked eye result reading style. PURPOSE: The purpose of this study was to establish and evaluate the novel numerical OFT-based parameter, so-called hemolysis area (HA), in screening thalassemia traits. MATERIALS AND METHODS: The portable spectrophotometer was invented capable of calculating the HA values. The HA values were then compared among 69, 156, and 19 blood samples having positive, negative, and suspicious 0.36% NaCl-based OFT results, respectively; 109 and 135 blood samples having mean corpuscular volume (MCV) ≤80 fL and >80 fL, respectively; and 138 and 106 blood samples having mean corpuscular hemoglobin (MCH) ≤27 pg and >27 pg, respectively. In addition, the HA values were compared in 166 blood samples having different globin gene genotypes. Finally, the HA cutoff value was determined by receiver operation curve (ROC) analysis. RESULTS: The HA values in samples having positive, suspicious, and negative 0.36% NaCl-based OFT were 33.3 ± 14.4, 42.9 ± 10.5, and 65.3 ± 13.4, respectively; in sample having MCV ≤80 fL and >80 fL were 43.1 ± 19.6 and 63.8 ± 14.5, respectively; and in samples having MCH ≤27 pg and >27 pg were 46.7 ± 20.1 and 64.8 ± 14.2, respectively. The HA values in normal, hemoglobin E, SEA-a thalassemia 1, and β-thalassemia traits were 67.1 ± 12.6, 36.4 ± 13.9, 20.2 ± 4.8, and 18.6 ± 1.1, respectively. All were significantly different. ROC analysis established 52.4 as the HA cutoff that had comparable effectiveness to the conventional screening tests. CONCLUSION: The new HA value was effective and could be an alternative choice for screening thalassemia traits.

Multiple-ancestry genome-wide association study identifies 27 loci associated with measures of hemolysis following blood storage

“…In addition, a number of novel GWA significant variations were found in metabolic enzymes (Hexokinase 1 -HK1; p<4.90E-11), stress kinases (MAPKAPK5; p<2.24E-13), ion channels (Piezo type mechanosensitive ion channel component 1; PIEZO1; p<4.04E-14), and other proteins, such as Myosin IXB (MYO9B; p<9.88E-15). Supporting the internal validity of these findings, many of these SNPs are in proteins known to cause RBC disorders such as spherocytosis (23), elliptocytosis (29), xerocytosis (30), and alpha-thalassemia (31).…”

“…We identified twenty loci that were genome-wide significant in all sample analysis (p<5x10 -8 ) for at least one of the hemolysis measures (Table 1). Many of the identified variants were concentrated in proteins known to cause human RBC disorders characterized RBC fragility such as dehydrated hereditary stomatocytosis (PIEZO1) (41,42), spherocytosis ANK1, SPTA1, SLC4A1 (23,43), ellipto-poikilocytosis (SPTA1) (44), xerocytosis (PIEZO1) (30), alpha-thalassemia (HBB) (31), and spontaneous and oxidant stress-induced hemolytic anemia (HK1 and G6PD) (16,45). Providing additional validity, many of the implicated SNPs have been associated with laboratory complete blood cell count measurements, such as reticulocyte counts (SPTA1 and PIEZO1) (46), and other complete blood count indices (G6PD) (47).…”

Laboratory Diagnosis of β-Thalassemia and HbE

“…The hemolysis area of normal individuals was 67.1 ± 12.6 units. In contrast, the hemolysis area of HbE carriers, β-thalassemia carriers were found to be 36.4 ± 13.9, 18.6 ± 1.1, respectively [54].…”

“…The β-thalassemia carriers all have the positive OFT results, while HbE carriers have either positive, negative, and suspicious OFT results (see the text for detail). Laboratory Diagnosis of β-Thalassemia and HbE DOI: http://dx.doi.org/10.5772/intechopen.90317cuvette[54]. To perform this test, 20 μL of EDTA blood is mixed with 5 mL of GSS in 12 × 75 polypropylene cuvette.…”

Alteration in the number, morphology, function, and metabolism of erythrocytes in high-altitude polycythemia

“…EOF is a measurement of the resistance of erythrocytes to hypotonic solutions and hemolysis, which primarily depends on the surface area/volume ratio in erythrocytes. Clinically, EOF can be applied to screening and evaluating certain hemolytic diseases, such as thalassemia, hereditary spherocytosis, and glucose-6-phosphate dehydrogenase deficiency ( Emilse et al, 2018 ; Tatu and Sweatman, 2018 ). Routine blood tests showed that the average erythrocyte volume in HAPC rats had increased, leading to a relative decrease in the surface area/volume ratio of erythrocytes, which may increase osmotic fragility.…”