Limited recombination events in merozoite surface protein-1 alleles of Plasmodium falciparum on islands

Abstract

Intragenic recombination is a principal mechanism for the generation of allelic variation in the merozoite surface protein-1 gene (Msp-1) of the human malaria parasite Plasmodium falciparum. In the present study, linkage disequilibrium between the 5′- and 3′-polymorphic sites was analyzed to determine the frequency of recombination events in Msp-1 in parasite populations on four islands in Vanuatu, the southwestern Pacific, where malaria transmission is moderate and comparable to other mesoendemic areas. Of 141 isolates, whose 5′-haplotypes (Msp-1 blocks 2–6) were determined by PCR-based typing, 138 were successfully sequenced for the 3′-polymorphism (block 17). A total of four distinct 5′-haplotypes and three distinct 3′-sequence types were identified with apparently different frequency distribution among islands. The number of 5′-haplotypes in each island was one to four, far smaller than in other previously studied geographic areas (ten to 21). Associations between the 5′- and 3′-polymorphisms (here termed Msp-1 gene types) were subjected to the R2 linkage disequilibrium test. The test revealed complete or very strong linkage disequilibrium in all four islands. Mixed infection was unusually rare (2.1%) and the mean number of Msp-1 alleles per person was nearly 1.0. The heterozygosity of the Msp-1 gene type calculated for each island (h=0.41–0.65) was significantly lower than that in other areas of comparable endemicity (h=0.81–0.89) (P<0.01). These results indicate that recombination events in Msp-1 would be extremely limited in Vanuatu, and stress that the frequency of recombination in Msp-1 is determined by not only the intensity of malaria transmission but the frequency of mixed clone infections, the mean number of clones per person and a repertoire of clones in a local area.