Molecular characterization of Plasmodium vivax clinical isolates in Pakistan and Iran using pvmsp-1, pvmsp-3alpha and pvcsp genes as molecular markers.
In this study, the diversity of Plasmodium vivax populations circulating in Pakistan and Iran has been investigated by using circumsporozoite protein (csp) and merozoite surface proteins 1 and 3alpha (msp-1 and msp-3alpha) genes as genetic markers. Infected P. vivax blood samples were collected from Pakistan (n=187) and Iran (n=150) during April to October 2008, and were analyzed using nested-PCR/RFLP and sequencing methods. Genotyping pvmsp-1 (variable block 5) revealed the presence of type 1, type 2 and recombinant type 3 allelic variants, with type 1 predominant, in both study areas. The sequence analysis of 33 P. vivax isolates from Pakistan and 30 from Iran identified 16 distinct alleles each, with one allele (R-8) from Iran which was not reported previously. Genotyping pvcsp gene also showed that VK210 type is predominant in both countries. Moreover, based on the size of amplified fragment of pvmsp-3alpha, three major types: type A (1800bp), type B (1500bp) and type C (1200bp), were distinguished among the examined isolates that type A was predominant among Pakistani (72.7%) and Iranian (77.3%) parasites. PCR/RFLP products of pvmsp-3alpha with HhaI and AluI have detected 40 and 39 distinct variants among Pakistani and Iranian examined isolates, respectively. Based on these three studied genes, the rate of combined multiple genotypes were 30% and 24.6% for Pakistani and Iranian P. vivax isolates, respectively. These results indicate an extensive diversity in the P. vivax populations in both studies.
Zakeri
Sedigheh
S
Malaria and Vector Research Group, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran. zakeris@yahoo.com
Raeisi
Ahmad
A
Afsharpad
Mandana
M
Kakar
Qutbuddin
Q
Ghasemi
Faezeh
F
Atta
Hoda
H
Zamani
Ghasem
G
Memon
Muhammad Suleiman
MS
Salehi
Masoud
M
Djadid
Navid Dinparast
ND
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics; D001483:Base Sequence; D005819:Genetic Markers; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D007492:Iran / Q000453:epidemiology; D016780:Malaria, Vivax / Q000453:epidemiology / Q000469:parasitology; D020066:Merozoite Surface Protein 1 / Q000235:genetics*; D008969:Molecular Sequence Data; D010154:Pakistan / Q000453:epidemiology; D010966:Plasmodium vivax / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D015800:Protozoan Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA
D000953:Antigens, Protozoan; D005819:Genetic Markers; D020066:Merozoite Surface Protein 1; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan