EE score: an index for simple differentiation of homozygous hemoglobin E and hemoglobin E-β0-thalassemia

Abstract

Abstract Background: The objective of the study was to describe a formula based on hemoglobin (Hb)A2 and HbF levels for differentiation of homozygous HbE and HbE-β-thalassemia. Methods: A total of 1256 subjects suspected for homozygous HbE or HbE-β0-thalassemia were recruited at the ongoing thalassemia screening program at Khon Kaen University, Thailand. Hb analysis was done using capillary electrophoresis. Genotyping was based on DNA analysis. An arbitrary formula based on HbA2 and HbF was developed statistically for differentiation of the two conditions. Validation was carried out prospectively on another 139 subjects encountered at routine laboratory. Results: Among 1256 subjects, Hb and DNA analyses identified cases with homozygous HbE (n=1076, 85.7%), HbE-β0-thalassemia (n=140, 11.1%), HbE-δβ0-thalassemia (n=30, 2.4%) and unknown HbE-related disorder (n=10, 0.8%). An inverse correlation between the amounts of HbA2 and HbF in HbE-β0-thalassemia was observed. With differences in the amounts of HbA2 and HbF between the groups, an arbitrary score (7.3 HbA2+HbF) was developed where score above 60 indicated HbE-β0-thalassemia. Application of this score on another 139 subjects showed accurate prediction of HbE-β0-thalassemia with 100% sensitivity, 96.5% specificity, 85.7% positive predictive value and 100% negative predictive value. Successful application onto couples at risk was demonstrated. Conclusions: An established score should prove useful in the differentiation of homozygous HbE and HbE-β0-thalassemia in routine setting and lead to a significant reduction in number of referring cases for molecular testing.