ABSTRACT
Background & objectives: β-thalassaemia is a genetic disorder and an important health problem around the world. Quantitative haemoglobin A2 (HbA2) levels are used for the diagnosis of β-thalassaemia. The conventional methods are high performance liquid chromatography (HPLC), electrophoresis, and microcolumn chromatography techniques. We established a fast protein liquid chromatography (FPLC) method, to measure quantitatively of HbA2 levels, and compared its efficacy with conventional methods. Methods: The FPLC method, using a DEAE Sepharose, Hi Trap anion-exchange column chromatography technique was set up for HbA2 measurement. In this study, 220 blood samples were screened for haemoglobin type by FPLC technique and also using HPLC, microcolumn chromatography and electrophoresis. Results: The FPLC results were highly correlated (r = 0.985, P<0.001) with those of HPLC for quantification of HbA2 as well as cellulose acetate electrophoresis (r = 0.977) and microcolumn chromatography (r = 0.980). The FPLC method showed 100 per cent sensitivity and specificity, positive and negative predictive value for β-thalassaemia diagnosis. In addition, the FPLC method was simple, rapid, low cost and reproducible. The HbA2/E range of FPLC for β-thalassaemia was 6-10 per cent, HbE trait was 10-40 per cent, β-thalassaemia/HbE was 40-60 per cent and homozygous HbE was more than 60 per cent. Interpretation & conclusions: Our findings suggested that FPLC method could be used as a cost-effective method for routine β-thalassaemia diagnosis.
Subject(s)
Adult , Chromatography, Ion Exchange/economics , Chromatography, Ion Exchange/methods , Chromatography, Ion Exchange/standards , Chromatography, Liquid/economics , Chromatography, Liquid/methods , Chromatography, Liquid/standards , Cost-Benefit Analysis , Electrophoresis/economics , Electrophoresis/methods , Electrophoresis/standards , Fetal Hemoglobin/analysis , Fetal Hemoglobin/isolation & purification , Hemoglobin A2/analysis , Hemoglobin A2/isolation & purification , Hemoglobin E/analysis , Hemoglobin E/isolation & purification , Hemoglobins/analysis , Hemoglobins/isolation & purification , Humans , Mass Screening/economics , Mass Screening/methods , Mass Screening/standards , Predictive Value of Tests , Sensitivity and Specificity , beta-Thalassemia/diagnosisABSTRACT
A sensitive,specific, and rapid method for the detection of carbohydrate-protein interactions is demonstrated by fluorophore-assisted carbohydrate electrophoresis (FACE). The procedure is simple and the cost is low. The advantage of this method is that carbohydrate-protein interactions can be easily displayed by FACE, and the carbohydrates do not need to be purified.