Evaluation of a new Sebia kit for analysis of hemoglobin fractions and variants on the Capillarys® system

Abstract

We evaluated the analytical performances of the new Sebia kit for quantification of hemoglobin fractions (HbA, HbF and HbA2) and structural hemoglobin variants on the Capillarys system. This automated capillary zone electrophoresis method uses an alkaline buffer with silica capillaries and spectrophotometric detection. Specimen stability was evaluated during 1 month. The reproducibility of migration and the imprecision of quantification were also investigated. Comparison with the Beckman P/ACE system was performed on 202 samples. A total of 131 subjects without any hematological abnormality were analyzed to establish the HbA2 reference ranges based on our local population. Quantification of the Hb fractions and variants exhibited excellent stability for 4 weeks of storage at 4 degrees C, with CVs < 0.3%. The imprecision of the migration normalized to that of HbA2 for all hemoglobins tested (fractions and variants) was low, with a CV of < 2.5%. At physiological and pathological levels, total imprecision ranged from 1.9% to 4.6% for HbA2, from 0.6% to 9.7% for HbF, and from 0.6% to 1% for HbS. Statistical analysis revealed a small proportional negative bias for HbA2 (-8.6%). Small systematic bias (-0.2%) and proportional bias (-28%) were observed for HbF. No statistically significant difference was found for HbS. The reference range for HbA2 was 2.1-3.2%. The Capillarys system is a fully automated and accurate system that gives high-resolution performance and displays appropriate characteristics for use as a routine method for the diagnosis of thalassemias and hemoglobinopathies.

Neonatal haemoglobinopathy screening in Belgium

“…New methods applicable to screening are now available (ie, capillary electrophoresis, tandem mass spectrometry) or are under development (ie, immunoassay) 9 – 11. A capillary electrophoresis system (Sebia Benelux, Brussels, Belgium) is under evaluation and we will soon evaluate an immunoassay (Perkin Elmer, Zaventem, Belgium).…”

Separation of haemoglobin HbE and HbA2 by the fully automated, high‐pressure liquid chromatography Tosoh HLC‐723 G7 analyzer

“…As a differentiation of HbE with either HbC‐Harlem or HbO‐Arab can be established by using appropriate tests, the problem remains with HbA 2 , as above‐mentioned. In a recent paper a capillary zone electrophoresis (CZE) procedure based on a commercial kit was demonstrated to clearly separate HbA 2 from HbE (Louahabi et al. , 2006).…”

Detection of &lt;i&gt;β&lt;/i&gt;-Hemoglobin Gene and Sickle Cell Disorder from Umbilical Cord Blood

“…Alkaline and acid hemoglobin electrophoresis methods are the most traditional and commonly used method to identify and investigate normal and mutant HbS [9] [10]. Moreover, hemoglobin fraction analysis of different hemoglobin variants by cation-exchange HPLC is another option to study hemoglobinopathies and other related disorders [10] [11]. Though these sophisticated techniques are being used in the developed countries, however, it is difficult to apply these advanced methods in low-income countries like Bangladesh for detection of hemoglobin disorders.…”