Genotyping and Phylogenetic Analysis of Plasmodium vivax Circumsporozoite Protein (PvCSP) Gene of Clinical Isolates in South-Eastern Iran
Abstract
Background: Circumsporozoite protein (CSP) is one of the most important surface sporozoite antigens in malaria, recently considered as a candidate for vaccination. Considering the importance of CSP, this study was conducted to investigate the polymorphism and genetic diversity of Plasmodium vivax Circumsporozoite Protein (Pvcsp) in the southeastern region of Iran during 2015-2016.
Methods: To investigate polymorphism and genetic diversity, 20 blood samples were collected from patients with P. vivax, then DNA was extracted and amplified using partial sequence of CSP gene. Polymerase chain reaction (PCR) products were sequenced and compared to sequences from genomic databases using BLAST. Genetic evaluation and phylogenic analysis were performed using MEGA7 and DnaSP5 software’s on 38 sequences include 20 sequences of our study and 18 sequences of Gene Bank.
Results: Eleven isolates were VK210 genotype and 9 isolates contained VK247. The result of variable segregation nucleotide site indicated that the differentiation of sequences in CSP were 25.67% in our 20 samples which are less than the 38 samples with a value of 26.67%. Comparing the ratio of dN/dS regions in the CSP gene indicates that the CSP varies more synonymously and amino acid has lower variation. Out of 38 samples, 35 unique haplotypes were identified based on 1042 nucleotide sequences in CSP, showing a variation percentage of 99.4%.
Conclusion: The Tajima D analyses showed that CSP gene in P. vivax had a positive number in the total analyzed sequences, which means that the P. vivax mutations are in order to select positive evolution.
Committee, W.M.P.A. and S.m.w. int (2016).Malaria Policy Advisory Committee to the WHO: conclusions and recommendations of eighth biannual meeting (September 2015). Springer.
Drexler, A.L., Y. Vodovotz, and S. Luckhart (2008). Plasmodium development in the mosquito: biology bottlenecks and opportunities for mathematical modeling. Trends in parasitology. 24(8): p. 333-336.
Hemami, M.R., et al (2013). Malaria elimination in iran, importance and challenges. International journal of preventive medicine.4(1) p. 88.
Mueller, A.-K., et al (2014). Invasion of mosquito salivary glands by malaria parasites: prerequisites and defense strategies. International journal for parasitology. 40(11): p. 1229-1235.
Cerami, C., et al (1992). The basolateral domain of the hepatocyte plasma membrane bears receptors for the circumsporozoite protein of Plasmodium falciparum sporozoites. Cell. 70(6): p. 1021-1033.
Coppi, A., et al (2011). The malaria circumsporozoite protein has two functional domains, each with distinct roles as sporozoites journey from mosquito to mammalian host. Journal of Experimental Medicine. 208(2): p. 341-356.
Goldman, I.F., et al (1993). Circumsporozoite protein gene of Plasmodium simium, a Plasmodium vivax-like monkey malaria parasite. Molecular and biochemical parasitology. 57(1): p. 177-180.
Kosaisavee, V., I. Hastings, and A. Craig (2011). The genetic polymorphism of Plasmodium vivax genes in endemic regions of Thailand. Asian Pacific journal of tropical medicine. 4(12): p. 931-936.
Imwong, M., et al (2005). Practical PCR genotyping protocols for Plasmodium vivax using Pvcs and Pvmsp1. Malaria Journal. 4(1): p. 20.
Kumar, S., G (2016). Stecher, and K. Tamura, MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular biology and evolution. 33(7): p. 1870-1874.
Saitou, N. and M. Nei (1987). The neighbor-joining method. a new method for reconstructing phylogenetic trees. Molecular biology and evolution, 1987. 4(4): p. 406-425.
Felsenstein, J (1985). Confidence limits on phylogenies an approach using the bootstrap. Evolution. 39(4): p. 783-791.
Kimura, M (1980)., A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of molecular evolution. 16(2): p. 111-120.
Rosenberg, R., et al (1989). Circumsporozoite protein heterogeneity in the human malaria parasite Plasmodium vivax. Science. 245(4921): p. 973-976.
Hedrick, P. (2011). Genetics of populations.: Jones & Bartlett Learning.
Howie, R.T., et al (2012). Mixed molecular and atomic phase of dense hydrogen. Physical Review Letters. 108(12): p. 125501.
Rootsi, S., et al (2004). Phylogeography of Y-chromosome haplogroup I reveals distinct domains of prehistoric gene flow in Europe. The American Journal of Human Genetics. 75(1): p. 128-137.
Singh, A.P., et al (2007). Plasmodium circumsporozoite protein promotes the development of the liver stages of the parasite. Cell. 131(3): p. 492-504.
Culleton, R., et al (2009). Evidence for the transmission of Plasmodium vivax in the Republic of the Congo, West Central Africa. The Journal of infectious diseases. 200(9): p. 1465-1469.
Hughes, A.L (2004). The evolution of amino acid repeat arrays in Plasmodium and other organisms. Journal of molecular evolution. 59(4): p. 528-535.
Dias, S., et al (2013). Population genetic structure of the Plasmodium vivax circumsporozoite protein (Pvcsp) in Sri Lanka. Gene. 518(2): p. 381-387.
Machado, R.L. and M.M. Póvoa (2000). Distribution of Plasmodium vivax variants (VK210, VK247 and P. vivax-like) in three endemic areas of the Amazon region of Brazil and their correlation with chloroquine treatment. Transactions of the Royal Society of Tropical Medicine and Hygiene. 94(4):p. 377-381.
Zakeri, S., et al (2006). Circumsporozoite protein gene diversity among temperate and tropical Plasmodium vivax isolates from Iran. Tropical Medicine & International Health. 11(5): p. 729-737.
Hernández-Martínez, M.Á., et al (2011). Antigenic diversity of the Plasmodium vivax circumsporozoite protein in parasite isolates of Western Colombia. The American journal of tropical medicine and hygiene. 84(2_Suppl): p. 51-57.
Lim, C.S., L. Tazi, and F.J. Ayala (2005). Plasmodium vivax: recent world expansion and genetic identity to Plasmodium simium. Proceedings of the National Academy of Sciences. 102(43): p. 15523-15528.
Mann, V.H., et al (1994). Sequence variation in the circumsporozoite protein gene of Plasmodium vivax appears to be regionally biased. Molecular and biochemical parasitology. 68(1): p. 45-52.
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Issue | Vol 49 No 5 (2020) | |
Section | Original Article(s) | |
Keywords | ||
Plasmodium vivax Circumsporozoite protein Genotyping Phylogenetic analysis Iran |
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