Screening of haemoglobinopathies by high performance liquid chromatography and its molecular characterization using amplification refractory mutation system and direct DNA sequencing techniques among college students in central Gujarat

Abstract

Background: Haemoglobinopathies are inherited blood disorders that can have significant health implications and are common in certain populations. Aim and Objectives: To investigate the prevalence of haemoglobinopathies among college students in central Gujarat, India. Material and Methods: A combination of techniques were employed to identify haemoglobinopathies. Chromatograms were examined to identify variant haemoglobin based on characteristics like proportion, retention time, and peak features. Prior to DNA analysis, prevalent Indian thalassemia mutations, as well as HbS and HbE, were identified using the Amplification Refractory Mutation System (ARMS) Polymerase Chain Reaction (PCR) technique. DNA sequencing was performed at SN Gene Lab, Surat. Results: The findings revealed that HbS codon 6 (A→T) heterozygosity, Hb S Codon 6 (A→T) double heterozygosity, and HbE: codon 26 (G→ A) were relatively common in the population. Among carrier students screened for haemoglobino-pathies, IVS-1, nt5 (G→C), and Codon41/42 (-CTTT) were the most prevalent beta-thalassemia mutations in India. Codon 88 (C-T) and other less common mutations were also detected. Conclusion: The study underscores the importance of raising awareness and providing counselling for individuals affected by haemoglobinopathies. By utilizing High Performance Liquid Chromatography (HPLC) screening and molecular techniques for characterization, the study contributes to the understanding of mutation patterns in central Gujarat. This knowledge is crucial for the effective detection of mutations in screening programs and prenatal diagnostics, ultimately enhancing the quality of life for affected individuals and preventing further transmission of these disorders.

Keywords

• amplification refractory mutation system polymerase chain reaction
• dna sequencing
• haemoglobinopathies
• high performance liquid chromatograph