Genetic diversity of Plasmodium falciparum infections in mild and severe malaria of children from Kampala, Uganda.

Diversity in parasite virulence is one of the factors that contribute to the clinical outcome of malaria infections. The association between the severity of Plasmodium falciparum malaria and the number of distinct parasite populations infecting the host (multiplicity of infection) or polymorphism within any of the specific antigen genes was investigated. The study included 164 children presenting with mild and severe malaria from central Uganda where malaria is meso-endemic. The polymorphic regions of the circumsporozoite protein (csp), merozoite surface proteins 1 and 2 (msp1 and msp2), and glutamate-rich protein (glurp) were genotyped by polymerase chain reaction methods and fragment analysis by gel electrophoresis. In a subset of samples fragment analysis was also performed by fluorescent PCR genotyping followed by capillary electrophoresis. The multiplicity of infection (MOI), determined as the highest number of alleles detected within any of the four genetic loci, was significantly higher in severe than in mild malaria cases (mean 3.7 and 3.0, respectively, P=0.002). No particular genotype or allelic family of msp1 or msp2 was associated with severity of malaria, and nor did the genotyping method reveal any significant difference in MOI when only assessed by msp2 genotyping. Severity of malaria was not linked to the predominance of any particular msp1 or msp2 allelic types, independent of methods used for genotyping. Monitoring the dynamics of multiple clone infections in relation to disease outcome, host immune status and genetic factors will provide more insight into parasite virulence mechanisms.