Ambareen Hamid, Ayisha Imran, Sobia Ashraf, Nauman Aslam Malik.
Screening Significant Haemoglobin Disorders HPLC Vs Electrophoresis.
Pak J Med Health Sci Jan ;12(2):710-12.

Background: Haemoglobin, protein responsible of carriage of gases consists of hetrotetramers of two alpha and two beta globin chains with one haem molecule in the center of each chain. Mutations of the globin genes can result in either a quantitative decrease output from that gene or modify the amino acid sequences of the protein produced. Quantitative defects produce thalassemia syndrome. The qualitative alterations are referred to as Hb variants. Inherited disorders of haemoglobin are the commonest single gene disorder. Screening and accurate identification of hemoglobin (Hb) variants have become increasingly important in antenatal diagnosis and prevention of Hb disorders. 5 Two most widely used screening methods are haemoglobin electrophoresis and high performance liquid chromatography (HPLC). Electrophoresis is one of the most common meth ods for haemoglobinopathy screening. Although cost-effective, the technique is relatively laborious, requiring manual sample preparation. High-performance liquid chromatography (HPLC), is a method that allows the detection of abnormal Hemoglobins quickly and precisely, using a small sample amount. I Aim: To compare the results of HPLC and Hemoglobin Electrophoresis technique for screening of clinically significant hemoglobin disorders. Methodology: This cross sectional comparative study was conducted at Chughtais Lahore Lab from August 2015 to October 2015. 100 consecutive patients of suspected haemoglobinopathy were included. Peripheral blood was collected in EDTA anticoagulant. Haemoglobin electrophoresis was done using cellulose acetate membrane at pH 8.4-8.6 on Interlab Genu electrophoresis system and Elfolab software version 7.03 was utilized for the calculation of results. The cases with co-migration on alkaline electrophoresis were confirmed on acid electrophoresis. All the samples were run in parallel on Biorad D10 HPLC analyzer using HbA2 and HbF programme. Results: A total of hundred cases were enrolled in study. Testing the samples yielded that 50(50%) had normal haemoglobin electrophoretic pattern. 30( 30%) had haemoglobin A 2 levels more than 3.5% (Thalasaemia trait) Thalasaemia major was detected in 13% of patients. Both the screening methods produce comparative results in detecting haemoglobinopathies. Conclusion: Our study denotes that the results obtained from the two techniques are comparable in screening significant haemoglobin disorders . There is considerable difference in cost, making screening by electrophoresis more affordable for economically burdened affected families

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