SCREENING AND MOLECULAR CHARACTERIZATION OF HAEMOGLOBINOPATHIES USING ARMS (AMPLIFICATION REFRACTORY MUTATION SYSTEM) AND DIRECT DNA SEQUENCING TECHNIQUES AMONG YOUNG IN CENTRAL GUJARAT WESTERN INDIA

Abstract

Haemoglobinopathies is consider the most common inherited disorders in human and results from genetic mutation in . one or more genes The present study was aimed to characterize the β-thalassemia mutation and haemoglobin variant in youth by ARMS PCR which is an uncomplicated and convenient method for identification of five common mutation from central Gujarat, western India region. This Study included 44 randomly selected haemoglobinopathies carrier student's sample of Anand People's Medicare Society (APMS), Anand for DNA analysis by ARMS PCR from March 2021-April 2021. Identification of five common Indian β thalassemia mutations along with Hb S and HB E were carried out by ARMS PCR method and δβ- thalassemia mutation was characterized by GAP-PCR. The samples which remain uncharacterized were sent to S N gene lab, Surat for DNA sequencing. In our study the most common mutation among five common mutations characterized was IVS-1, nt5 (G→C) in 22 (50%), followed by Codon41/42 (-CTTT) in 5 (11.3%). IVS-1, nt1 (G→T), Codon8/9 (+G) and 619bp del mutation was not identified in any carrier students screened for haemoglobinopathies. Other than these five common mutation Codon -88 (C→T) (2.27%) and Codon 30 (G→A) (2.27%) are also detected. The prevalence of haemoglobinopathies with respect to communities, reflects that SC/ST/OBC are at the highest risk with 50%. Communities like Rajput (22.7%), Patel (18.1%), Brahmin (6.8%) and Muslim (2.2%) are also showing prevalence. The study has included mutation in different communities reflects characterization of mutation of central Gujarat, western India which is significant for rapid and convenient identification of mutation while conducting screening programs and prenatal diagnosis. Rare mutations which are not recognized, need further confirmation for carrier detection followed by prenatal diagnosis.