Abstract
BackgroundPlasmodium vivax reticulocyte binding protein-2 (PvRBP-2) is a promising candidate for development of vaccine against parasite. DNA sequence polymorphism in pvrbp-2 which may hamper the vaccine development program has been identified in laboratory strains. Therefore, unraveling genetic polymorphism in pvrbp-2 from field isolates is a prerequisite for success in vaccine development. This study was designed with a primary aim to uncover genetic polymorphism in pvrbp-2 among P. vivax field isolates.ResultsUsing virtual restriction mapping of pvrbp-2 sequences, two restriction enzymes (AluI and ApoI) were selected for the development of pvrbp-2 as a PCR-RFLP marker. Restriction fragment length polymorphism (RFLP) analysis revealed a high degree of genetic polymorphism in the pvrbp-2 gene among field isolates of P. vivax. ApoI-RFLP was found to be more efficient in identifying the extent of genetic polymorphism in pvrbp-2 compared to AluI-RFLP. Combined genotyping/haplotyping of RFLP pattern revealed a total of 36 distinct RFLP patterns among 83 P. vivax isolates analyzed. DNA sequence analysis also supports high degree of genetic polymorphism among field isolates of P. vivax. Pvrbp-2 PCR-RFLP method is able to distinguish multiple infection up to 16.86% and it revealed a low level of shared genetic pool between more than two populations.ConclusionThe study suggests that pvrbp-2 is highly polymorphic genetic marker which can be used for population genetic analyses. RFLP analysis suggests presence of nearly similar proportion of Sal-1 and Belem alleles in Indian P. vivax populations. The larger extent of genetic polymorphism identified from limited samples advocates to screen genetic polymorphism in pvrbp-2 from malaria endemic geographical regions and countries for designing pvrbp-2 based anti-malarial control measures.
Highlights
Plasmodium vivax reticulocyte binding protein-2 (PvRBP-2) is a promising candidate for development of vaccine against parasite
The sequence polymorphism reported in pvrbp-2 from four strains of P. vivax including Sal-1 and Belem [22] is supporting the extent of genetic polymorphism observed in pvrbp-2 in Indian isolates
The study suggests that pvrbp-2 is highly polymorphic genetic marker which can be used for population genetic analyses
Summary
Plasmodium vivax reticulocyte binding protein-2 (PvRBP-2) is a promising candidate for development of vaccine against parasite. DNA sequence polymorphism in pvrbp-2 which may hamper the vaccine development program has been identified in laboratory strains. Unraveling genetic polymorphism in pvrbp-2 from field isolates is a prerequisite for success in vaccine development. This study was designed with a primary aim to uncover genetic polymorphism in pvrbp-2 among P. vivax field isolates. Invasion of erythrocytes by malaria parasite is a complex and multi-step process. The major function of reticulocyte binding protein is seen during the initial steps of erythrocyte selection and invasion [17]. Evidence suggests that the PvRBPs form a complex at the apical pole of the merozoite and confer the reticulocyte-specificity of P. vivax blood-stage infections, suggesting the essential role of RBP-II in selection and identification of reticulocyte for invasion [17]. Two pvrbp-2 genes have been characterized from P. vivax and are shown to be a promising vaccine candidate [20]; up to 12 putative pvrbp genes have been identified in P. vivax genome so far [21]
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