A multiplex ARMS PCR approach to detection of common β-globin gene mutations

Background: beta-thalassaemia is a group of inherited single-gene disorders worldwide. Each ethnic population has its own common mutations. Heterogeneity of beta-thalassaemia mutations in multi-ethnic population of Surat, makes molecular diagnosis expensive and time consuming. Methods: Specific primers were used to differentiate four common mutations, IVS I-5 (G -> C), Codon 41/42 (-TCTT), 619-bp deletion and FS 8/9 (+G), by a simple PCR involving a multiplex amplification refractory mutation system. Results: Several high prevalence beta-Thalassemia trait groups constituted by Muslims, Patels, Sindhis, ModhBanias, and Mahayavanshi. Four most common mutations detected in them are IVS I-5 (G -> C), Codon 41/42 (-TCTT), 619-bp deletion and FS 8/9 (+G). We identified each of these beta-thalassemia mutations in multiplexed ARMS from positive control samples. Our multiplex-ARMS-PCR system was first standardized on positive DNA samples with above known four most common beta-thalassemia mutations, and these positive samples had been diagnosed with beta-thalassemia and also all these samples belonged to Surat ethnic groups. The system was subsequently tested on 110 blood samples from different ethnic backgrounds with unknown beta-thalassemia mutations which were in all specimens. Conclusion: The ARMS multiplex system was found reliable, cost effective, fast and most applicable for mutation screening of Thalassemia in Surat populations. (C) 2017 Elsevier Inc. All rights reserved.