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Background   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. 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.    Results  Using virtual restriction mapping of  pvrbp-2  sequences, two restriction enzymes ( Alu I and  Apo I) 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 .  Apo I-RFLP was found to be more efficient in identifying the extent of genetic polymorphism in  pvrbp-2  compared to  Alu I-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.    Conclusion  The 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.

Molecular analysis of reticulocyte binding protein-2 gene in Plasmodium vivax isolates from India