Original Article

Genetic Variation of MSP-1 Gene in Plasmodium vivax Isolated from Patients in Hormozgan Province, Iran using SSCP-PCR

Abstract

Background: The main goal of present study was to detect polymorphism in MSP-1 gene which is a major blood stage candidate for vaccine in Plasmodium vivax by Single Strand Conformational Poly- morphism-Polymerase Chain Reaction (SSCP-PCR).

Methods: During 2008 to 2010 fifty samples were collected from Iranian patients with P. vivax in Hormozgan Province, southern Iran. All of the samples were detected by microscopical examination. Amplification of MSP-1 gene was done by PCR after DNA extraction. Single strand DNAs due to using in SSCP, was electrophoresed on polyacrylamid- Bisacrylamid gel then banding patterns were revealed by silver-staining method. Sequencing as a typing method was performed for some isolates.

Results: All of the 50 isolates were positive microscopically. Totally 12 (24%) isolates showed 440 bp and 38 (76%) showed 500 bp in PCR assay. SSCP analysis revealed four banding patterns. Pattern I  (10/50), Pattern II (12/50), Pattern III (27/50), and Pattern IV (1/50). The results sequencing analysis of the MSP-1 gene in 19 isolates revealed diversity in nucleotides and amino acid in Iranian P. vivax isolates.

Conclusion: Our study confirms that the SSCP-PCR is a rapid method for detecting polymorphism in MSP-1 gene in P. vivax. The presence of different haplotypes in MSP-1 gene shows that several P. vivax strains exist in malaria endemic areas of Iran.

 

Yeshiwondim AK, Tekle AH, Dengela DO, Yohannes AM, Teklehaimanot A. Therapeutic efficacy of chloroquine and chloroquine plus primaquine for thetreatment of Plasmodium vivax in Ethiopia. Acta Trop. 2010;113(2):105-13.

Arnott A, Barry AE, Reeder JC. Understanding the population genetics of Plasmodium vivax is essential for malaria control and elimination. Malar J. 2012; 11:14.

Guerra CA, Howes RE, Patil AP, Gething PW, Van Boeckel TP, Temperley WH. The international limits and population at risk of Plasmodium vivax transmission in 2009. PLOS Neglected Trop Dise. 2010; 8:e744.

Baird JK. Chloroquine resistance in Plasmodium vivax. Antimicrobial AgentChemother. 2004; 48:4075-4083.

Heidari H, Keshavarz H, Shojaee S, Raeisi A, Dittrich S. In vivo susceptibility of Plasmodium vivax to chloroquine in Southeastern Iran. Iranian J Parasitol. 2012; (In press)

Cui L, Escalante AA, Imwong M, Snounou G. The genetic diversity of Plasmodium vivax populations. Terend Parasitol. 2003; 19: 220-226.

Udagama PV, Gamage-Mendis AC, David PH, Peiris JS, Perera KL, Mendis KN. Genetic complexity of Plasmodium vivax parasites in individual human infections analyzed with monoclonal antibodies against variant epitopes on a single parasite protein. Am J Trop Med Hyg. 1990; 42: 104-110.

Gibson HL, Tucker JE, Kaslow DC, Krettli AU, Collins WE, Kiefer MC. Structure and expression of the gene for Pv200, a major blood-stage surface antigen of Plasmodium vivax. Mol Bioch Parasitol.1992;50: 325-533.

Kolakovich KA, Ssengoba A, Wojcik K, Tsuboi T, Al-Yaman F, Alpers M. Plasmodium vivax: favored gene frequencies of the merozoite surface protein-1 and the multiplicity of infection in a malaria endemic region. Exp Parasitol. 1996; 83: 11-18.

Del Portillo HA, Longacre S, Khouri E, David PH. Primary structure of the merozoite surface antigen 1 of Plasmodium vivax reveals sequences conserved between different Plasmodium species. Proc Natd Acad Sci. 1991;88; 4030-4034.

Hayashi K. PCR-SSCP: a method for detection of mutations. Genetic Analysis. 1992;9:73-79.

Orita M, Iwahana H, Kanazawa H, Hayashi K, Sekiya T. Detection of polymorphisms of human DNA by gel electrophoresis as singlestrand conformation polymorphisms. Proc Natd Acad Sci. 1989; 86: 2766-2770.

Sekiya T. Detection of mutant sequences by single-strand conformation polymorphism analysis. Mutation Research.1993; 288:79-83.

Slabaugh MB, Huestis GM, Leonard J, Holloway JL, Rosato C, Hongtrakul V. Sequence-based genetic markers for genes and gene families: single-strand conformational polymorphisms for the fatty acid synthesis genes of Cuphea. TAG Theoretical and Applied Genetics.1997;14: 400-408.

Craig AA, Kain KC. Molecular analysis of strains of Plasmodium vivax from paired primary and relapse infections. JID. 1996;174; 373-379.

Brant J, Bassam BJ, Caetano-Anollis G, Gresshoff PM. Fast and sensitive silver staining of DNA in polyacrylamide gels. Analytical and Biochemistery.1991;196; 80-83.

Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol. 2007; 24: 1596–1599.

Gutierrez A, Vicini J, Patarroyo ME, Murillo LA, Patarroyo MA. Plasmodium vivax: polymorphism in the merozoite surface protein 1 gene from wild Colombian isolates. Exp Parasitol.2000;95: 215-219.

Mallika I, Sasithon P, Laurent R, Frank L, Sornchai L, Georges S. Practical PCR genotyping protocols for Plasmodium vivax using Pvcs and Pvmsp1. Malaria J. 2005;4:20.

Zakeri S, Mehrizi AA, Mamaghani S, Noorizadeh S, Snounou G, Djadid ND. Population structure analysis of Plasmodium vivax in areas of Iran with different malaria endemicity. Am J Med Hyg. 2006;74: 394-400.

Kim JR, Imwong M, Nandy A, Chotivanich K, Nontprasert A, Tonomsing N. Genetic diversity of Plasmodium vivax in Kolkata, Indian. Malaria J. 2006;5:71.

Farooq U, Malla N, Dubey ML. Polymorphism in merozoite surface protein-1 gene in north & northwestern Indian field isolates of Plasmodium vivax. Indian J Med Res. 2009; 130:736-741.

Tsuboi T, Kaneko O, Cao YM, Tachibana M, Yakushijin Y, Nagao T. A rapid genotyping method for the vivax malaria transmissionblocking vaccine candidates, Pvs25 and Pvs28. Parasitol Int. 2004; 53: 211-216.

Files
IssueVol 7 No 4 (2012) QRcode
SectionOriginal Article(s)
Keywords
Plasmodium vivax MSP-1 Polymorphism SSCP-PCR Iran

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Miahipour A, Keshavarz H, Heidari A, Raeisi A, Rezaeian M, Rezaie S. Genetic Variation of MSP-1 Gene in Plasmodium vivax Isolated from Patients in Hormozgan Province, Iran using SSCP-PCR. Iran J Parasitol. 2015;7(4):1-7.