Vol 15, Issue 7, 2022 Online - 2455-3891 Print - 0974-2441 THE INCIDENCE OF BETA-THALASSEMIA MINOR IN PREGNANT FEMALES BY MEASURING HBA2 THROUGH HIGH PERFORMANCE LIQUID CHROMATOGRAPHY ANITA CHAUDHARY 1 , NINDER KUMAR 1 *, RITU KUNDAL 2 , RAMESH KUMAR 1 , PREET KAMAL SIBIA 3 1 Department of Pathology, Government Medical College, Patiala, Punjab, India. 2 Department of Peadiatrics, Maharishi Markandeshwar Medical College, Solan, Himachal Pradesh, India. 3 Department of Obstetrics and Gynaecology, Government Medical College, Patiala, Punjab, India. Email: drninder@gmail.com Received: 04 April 2022, Revised and Accepted: 10 May 2022 ABSTRACT Objectives: The aim of the study was to study the incidence of thalassemia minor by determining HbA2 levels in pregnant females attending Obstetrics and Gynaecology OPD and HbA2 levels of husbands of positive females for beta-thalassemia trait. Methods: The prospective study spanning over 1½ years was conducted in the Department of Pathology in 1020 pregnant females who attended the Gynaecology and Obstetrics OPD of Government Medical College, Patiala for antenatal check-up. The pregnant females of any trimester without any specific sign and symptoms whose Hb level was <10 g/dL were screened in the study. Levels of HbA2 and HbF were determined by high performance liquid chromatography (HPLC) and the cases with raised HbA2 value above the cutoff limit (>3.5%) were labeled as BTT. Husbands of BTT positive females were also screened for the trait. Incidence of all these cases was calculated and analyzed statistically. Results: The majority of the females were in the age group of 21–30 years. In present study, we found that total 134 (13.1%) patients were having beta thalassemia trait. Husbands of all these positive patients were also screened for BTT and only 2 (1.49%) of them were found to be positive. Conclusion: HPLC has the advantage for screening and detection of various hemoglobinopathies by providing rapid and accurate results. HPLC can detect and measure HbF and HbA2 in a single system. Early diagnosis and management of thalassemia can help in reduction of burden on society as well as government. Keywords: Beta thalassemia, Hemoglobinopathies, HbA2, HbF, High performance liquid chromatography. INTRODUCTION Human hemoglobin consists of two pairs of globin chains with heme group attached to each of them. The individual globin chains synthesized in postnatal life are designated as α, β, γ, and δ. HbA has two α chains and two β chains (α2 β 2); HbF has two α chains and two γ chains (α2 γ 2) and HbA2 has two α chains and two δ chains (α2 δ2). Alpha chain synthesis is directed by two α genes, α 1 and α β, on chromosome 16, and β and δ chain synthesis by single β and δ genes on chromosome 11. γ chain synthesis is directed by two genes, G γ and A γ, which are also located on chromosome 11 [1]. HbF is the predominant hemoglobin of fetal life where as in children and adults, HbA is the major hemoglobin. HbA2 and HbF are found in small quantities in adult life (2–3.3% and 0.2–1.0%, respectively). The adult proportions of hemoglobin A, A2, and F are usually attained by 6–12 months of age [1]. The disorders of hemoglobin synthesis can be grouped into three main categories: 1. Due to structural variants of hemoglobin, such as HbS. 2. Due to failure of synthesis of one or more globin chains of hemoglobin at a normal rate, as in thalassemias. 3. Those due to failure to complete the normal neonatal switch from fetal hemoglobin to adult hemoglobin referred to as hereditary persistence of fetal hemoglobin (HPFH) [1]. Thalassemias refer to heterogeneous group of disorders in which there is decreased or absent synthesis of one or more polypeptide chains (α or β) as a result of missense/non-sense mutations (single- base substitutions) or frameshift mutations of the genes controlling the structure of the hemoglobin protein chains in one or both “allelic” globin genes causing decreased hemoglobin concentration, microcytosis, and anemia [2]. The two major categories are the α and β thalassemia while the others are rare forms. These subgroups have in common an imbalanced globin synthesis and the globin produced in excess is responsible for ineffective erythropoiesis and hemolysis. The thalassemias result from the effect of a number of different molecular defects leading to a variety of clinical and hematologic phenotypes. Functionally, thalassemia mutations that cause a complete absence of globin chain synthesis are called α0 or β0 thalassemias and that cause reduced rate of synthesis of globin chains are called α+ or β+ thalassemias [3]. Clinically, the thalassemia is classified according to their severity into major, intermediate, and minor forms [3]. Thalassemias are among the most common genetic disorders in the world and an estimated 1.5% of the worldwide population is carrier of β-thalassemia [4]. Thalassemia is mainly prevalent in South-east Asia and Mediterranean countries. Due to high rate of migration and inter racial marriage system, thalassemia is found all over the world in recent years. About 10% of the total world thalassemia patients belong to Indian subcontinent and among them 3–4% are carriers [5]. β thalassemia is the most common single gene disorder in India. More than 30 million people worldwide carry thalassemia causing defective gene. Carrier frequency varies from 3 to 17% in different populations [6]. Definite diagnosis of thalassemia can be made by a step-wise algorithmic approach, starting with a detailed clinical history, through hematologic evaluation complete blood count (CBC)], reticulocyte count, red blood cell (RBC) morphology, Naked Eye Single Tube Red Cell Osmotic Fragility Test (NESTROFT), protein based analytic methods [Hb electrophoresis or isoelectric focusing (IEF), cation exchange high performance liquid chromatography (HPLC), reversed phase HPLC] © 2022 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/ licenses/by/4.0/) DOI: http://dx.doi.org/10.22159/ajpcr.2022v15i7.44840. Journal homepage: https://innovareacademics.in/journals/index.php/ajpcr Research Article