title: Molecular genetic analysis of Plasmodium vivax isolates from Eastern and Central Sudan using pvcsp and pvmsp-3α genes as molecular markers.
abstract: In Sudan, Plasmodium vivax accounts for approximately 5-10% of malaria cases. This study was carried out to determine the genetic diversity of P. vivax population from Sudan by analyzing the polymorphism of P. vivax csp (pvcsp) and pvmsp-3α genes. Blood samples (n=76) were taken from suspected malaria cases from 2012-2013 in three health centers of Eastern and Central Sudan. Parasite detection was performed by microscopy and molecular techniques, and genotyping of both genes was performed by PCR-RFLP followed by DNA sequence for only pvcsp gene (n=30). Based on microscopy analysis, 76 (%100) patients were infected with P. vivax, whereas nested-PCR results showed that 86.8% (n=66), 3.9% (n=3), and 3.9% (n=3) of tested samples had P. vivax as well as Plasmodium falciparum mono- and mixed infections, respectively. Four out of 76 samples had no results in molecular diagnosis. All sequenced samples were found to be of VK210 (100%) genotype with six distinct amino acid haplotypes, and 210A (66.7%) was the most prevalent haplotype. The Sudanese isolates displayed variations in the peptide repeat motifs (PRMs) ranging from 17 to 19 with GDRADGQPA (PRM1), GDRAAGQPA (PRM2) and DDRAAGQPA (PRM3). Also, 54 polymorphic sites with 56 mutations were found in repeat and post-repeat regions of the pvcsp and the overall nucleotide diversity (π) was 0.02149±0.00539. A negative value of dN-dS (-0.0344) was found that suggested a significant purifying selection of Sudanese pvcsp, (Z test, P<0.05). Regarding pvmsp-3α, three types were detected: types A (94.6%, 52/55), type C (3.6%, 2/55), and type B (1.8%, 1/55). No multiclonal infections were detected, and RFLP analysis identified 13 (Hha I, A1-A11, B1, and C1) and 16 (Alu I, A1-A14, B1, and C1) distinct allelic forms. In conclusion, genetic investigation among Sudanese P. vivax isolates indicated that this antigen showed limited antigenic diversity.
authors.affiliation: Department of Parasitology, Blue Nile National Institute for Communicable Diseases, University of Gezira, P.O. Box 20, Wad Medani, Sudan; Department of Parasitology, Faculty of Medical Laboratory Sciences, University of Gezira, P.O. Box 20, Wad Medani, Sudan.
authors.forename: Albadawi Abdelbagi
authors.initials: AA
authors.lastname: Talha
authors.affiliation: Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Pasteur Avenue, P.O. Box 1316943551, Tehran, Iran.
authors.forename: Sekineh
authors.initials: S
authors.lastname: Pirahmadi
authors.affiliation: Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Pasteur Avenue, P.O. Box 1316943551, Tehran, Iran.
authors.forename: Akram Abouie
authors.initials: AA
authors.lastname: Mehrizi
authors.affiliation: Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Pasteur Avenue, P.O. Box 1316943551, Tehran, Iran.
authors.forename: Navid Dinparast
authors.initials: ND
authors.lastname: Djadid
authors.affiliation: Department of Parasitology, Blue Nile National Institute for Communicable Diseases, University of Gezira, P.O. Box 20, Wad Medani, Sudan; Department of Parasitology, Faculty of Medical Laboratory Sciences, University of Gezira, P.O. Box 20, Wad Medani, Sudan.
authors.forename: Bakri Y M
authors.initials: BY
authors.lastname: Nour
authors.affiliation: Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Pasteur Avenue, P.O. Box 1316943551, Tehran, Iran. Electronic address: zakeris@yahoo.com.
authors.forename: Sedigheh
authors.initials: S
authors.lastname: Zakeri
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000953:Antigens, Protozoan / Q000235:genetics*; D002648:Child; D002675:Child, Preschool; D005260:Female; D017125:Genes, Protozoan / Q000235:genetics; D005819:Genetic Markers / Q000235:genetics; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D016780:Malaria, Vivax / Q000453:epidemiology / Q000469:parasitology; D008297:Male; D008875:Middle Aged; D010966:Plasmodium vivax / Q000235:genetics*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D015800:Protozoan Proteins / Q000235:genetics*; D013397:Sudan; D055815:Young Adult
chemical_list: D000953:Antigens, Protozoan; D005819:Genetic Markers; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan; C436736:merozoite surface protein 3gamma
keywords: Genetic analysis; Malaria; Plasmodium vivax; Sudan; csp; msp-3α