Studies on polymorphism in exon 5 of the GYPB gene in malaria positive patients in Punjab, Pakistan

Abstract

Background: Malaria, caused by the Plasmodium parasite, continues to be a major worldwide health problem, particularly in endemic countries, such as Pakistan. The purpose of this study is to examine the polymorphism in exon 5 of the GYPB gene in individuals who tested positive for malaria in Khushab, Punjab, Pakistan. Methodology: Cross-sectional study involved the collection of blood samples from malaria positive patients for the extraction of genomic DNA by the phenol-chloroform method. Primers were designed, optimized, and used for PCR amplification of Exon 5 of GYPB gene. PCR products were confirmed through agarose gel electrophoresis, succeeded by Sanger sequencing and mutation analysis using bioinformatics tools. Results: Among 35 malaria-positive cases, 52.14 % had P. vivax infection , 28.57% had P. falciparum infection , and 14.29 % mixed infections. Sequencing revealed polymorphisms and mutations in the exonic, intronic, and 3’UTR regions of the GYPB gene. Three of mutations were already reported and 5 are the novel ones. Samples ME5.32 (c.272C>A) and ME5.18 (g.41355T>C) displayed reported mutation, resulting in an alteration of an amino acid (A91E). Novel mutations were identified in samples ME5.32, ME5.24, ME5.21, characterized by g.144462A>G, g.144507C>T, and g.144498T>C, respectively. ME5.18 demonstrated novel polymorphisms and mutations: g.144522_144523insG, g.144498T>C, and g.41355T>C. Conclusion: The study revealed unique genetic variations within the GYPB gene, among malaria-infected individuals in Khushab. Pakistan. These findings highlight the need for further widescale investigations to understand the genetic landscape of malaria in the Pakistani population and its significance for disease susceptibility and treatment.