Evaluation of four different HPLC devices for hemoglobinopathy screening

Müjgan Ercan Karadağ,Gülsüm Abuşoğlu,Fatma Meriç Yılmaz,Saadet Kader,Emiş Deniz Akbulut,Esin Avcı,Muhittin Serdar,Esra Fırat Oğuz

doi:10.1515/tjb-2019-0484

Abstract

Abstract Objective Hemoglobinopathies are a common public health problem in Turkey. In the screening of these disorders in population, cation-exchange high performance liquid chromatography (HPLC) is accepted as the gold standard method. In this study, the aim was to assess four different HPLC devices used in hemoglobinopathy screening. Materials and methods A total of 58 blood samples were analyzed with four different HPLC methods (Bio-Rad variant II, Agilent 1100, Tosoh G8 and Trinity Ultra2 trademarks). Results The comparison study demonstrated a good correlation between the results of each HPLC analyzer and the reference value obtained by averaging all the HbA2 results belonging to the methods tested in the study [ (Tosoh G8 (r=0.988), Bio-Rad variant II (r=0.993), Agilent 1100 (r=0.98) and Trinity Ultra2 (r=0.992) ]. HbA2 determination in the presence of HbE was interfered in both Bio-Rad variant II and Tosoh G8. Conclusion The analyzers were found to have compatible HbA2 results but with accompanying different degrees of proportional and systematic biases. HPLC analyzers may be affected by different hemoglobin variants at different HbA2 concentrations, which is an important point to take into consideration during the evaluation of HbA2 results in thalassemia screening.

Highlights

Hemoglobinopathies, a common public health problem in Turkey, comprise a diverse group of diseases including thalassemia and hemoglobin variants
high performance liquid chromatography (HPLC) analyzers may be affected by different hemoglobin variants at different HbA2 concentrations, which is an important point to take into consideration during the evaluation of HbA2 results in thalassemia screening
Fight against hemoglobinopathies must essentially include the detection of thalassemia carriers, hemoglobin variants expected to present with severe clinical manifestations and clinically silent hemoglobin variants [3, 4]

Summary

Introduction

Hemoglobinopathies, a common public health problem in Turkey, comprise a diverse group of diseases including thalassemia and hemoglobin variants. When the prevalence of carriers is considered, 300– 400 affected newborns are expected per year and this causes significant emotional and economic harm to public health. Fight against hemoglobinopathies must essentially include the detection of thalassemia carriers, hemoglobin variants expected to present with severe clinical manifestations and clinically silent hemoglobin variants [3, 4]. Quantitative detection of HbA2 and HbF levels and usage of red cell indices are the main methodologies in hemoglobinopathy screening [5]. For being fully automated and allowing both qualitative and quantitative analysis of Hb fractions, cation exchange HPLC is accepted as the gold standard for population screening of hemoglobinopathies [5, 8]

Methods

Results

Discussion

Conclusion