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

Prajapati SK, Kumari P, Singh OP - BMC Microbiol. (2012)

Bottom Line: ApoI-RFLP was found to be more efficient in identifying the extent of genetic polymorphism in pvrbp-2 compared to AluI-RFLP.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular Biology Division, National Institute of Malaria Research, Sector 8, Dwarka, Delhi, India. surendramrc@gmail.com

ABSTRACT

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 (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.

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.

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Number of unique and shared pvrbp-2 genotypes among study sites.
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Figure 5: Number of unique and shared pvrbp-2 genotypes among study sites.

Mentions: Genetic polymorphism was observed among all geographical regions of the Indian subcontinent. The total number of genotypes observed in Delhi, Nadiad, Panna, Rourkela, Chennai and Kamrup were 11, 14, 12, 7, 7 and 4 respectively. In every geographical region, genotypes were observed to be unique (local polymorphism) and shared in varied proportion (Figure 5). Allelic analysis shows a limited fraction of genotypes were observed to have been shared within 2–3 populations. Only a single genotype (14) was observed in five geographical regions, however, none of the shared genotypes was observed between six geographical regions. This study suggests a diverse pool of pvrbp-2 repertoire in all geographical regions. This study also uncovered many unique pvrbp-2 genotypes to exist among geographical regions.


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

Prajapati SK, Kumari P, Singh OP - BMC Microbiol. (2012)

Number of unique and shared pvrbp-2 genotypes among study sites.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3539960&req=5

Figure 5: Number of unique and shared pvrbp-2 genotypes among study sites.
Mentions: Genetic polymorphism was observed among all geographical regions of the Indian subcontinent. The total number of genotypes observed in Delhi, Nadiad, Panna, Rourkela, Chennai and Kamrup were 11, 14, 12, 7, 7 and 4 respectively. In every geographical region, genotypes were observed to be unique (local polymorphism) and shared in varied proportion (Figure 5). Allelic analysis shows a limited fraction of genotypes were observed to have been shared within 2–3 populations. Only a single genotype (14) was observed in five geographical regions, however, none of the shared genotypes was observed between six geographical regions. This study suggests a diverse pool of pvrbp-2 repertoire in all geographical regions. This study also uncovered many unique pvrbp-2 genotypes to exist among geographical regions.

Bottom Line: ApoI-RFLP was found to be more efficient in identifying the extent of genetic polymorphism in pvrbp-2 compared to AluI-RFLP.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular Biology Division, National Institute of Malaria Research, Sector 8, Dwarka, Delhi, India. surendramrc@gmail.com

ABSTRACT

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 (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.

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.

Show MeSH
Related in: MedlinePlus