Abstract
The aim of this study was to determine the genetic diversity of Plasmodium falciparum by analyzing the polymorphism of the msp-1 and msp-2 genes and the multiplicity of infection in children with uncomplicated malaria in southern Benin. Blood samples of children with fever or history of fever with thick smear positive P. falciparum were collected on filter paper. After extraction of DNA by Chelex®, the samples underwent nested PCR. 93 isolates from children were genotyped. For the msp-1 gene, the K1 and R033 sequences were the most represented in the study population with 85.2% and 83% prevalence, respectively. Regarding the msp-2 gene, the FC27 family was more highly represented with 99% prevalence against 81.5% for 3D7. Mixed infections accounted for 80.4% of the samples. Twenty-five alleles were identified for msp-1 and 28 for msp-2. Fourteen and ten alleles belonged to the K1 (100–500 bp) and MAD20 (100–500 bp) families, respectively. The RO33 sequence did not show any polymorphism, with only one variant (160 bp) detected. The msp-2 gene was present as 16 FC27 family fragments (250–800 bp) and 12 of the 3D7 family (350–700 bp). The multiplicity of infection was estimated at 3.8 for msp-1 and 3.9 for msp-2 with 77 (87.5%) and 84 (91.3%) samples harboring more than one parasite genotype for msp-1 and msp-2, respectively. The multiplicity of infection (MOI) was influenced neither by age nor by parasite density. This study shows a significant diversity of P. falciparum in southern Benin with an MOI unaffected by age or by parasite density.
Highlights
IntroductionSample collection and laboratory proceduresIn Benin, P falciparum infection is among the first diseases and is responsible for 36% of deaths among children under 5 years (unpublished data from Ministry of Health)
Sample collection and laboratory proceduresIn Benin, P falciparum infection is among the first diseases and is responsible for 36% of deaths among children under 5 years
This study aimed to characterize the allelic polymorphism of msp-1 and msp-2 and determined the multiplicity of infection in P. falciparum isolates collected from children with uncomplicated malaria living in the southern Benin
Summary
Sample collection and laboratory proceduresIn Benin, P falciparum infection is among the first diseases and is responsible for 36% of deaths among children under 5 years (unpublished data from Ministry of Health). One of the limitations to the development of this vaccine against P. falciparum is the extensive genetic diversity in parasite populations limiting the efficacy of acquired protective immunity to malaria [6]. Asexual blood stage antigens, such as merozoite surface protein-1 (msp-1) and merozoite surface protein-2 (msp-2), are considered prime candidates for the development of a malaria vaccine and are suitable markers for the identification of genetically distinct P. falciparum parasite sub-populations [5]. Following scale up of malaria control interventions (massive deployment of insecticide-treated nets and free treatment with ACT) in the country, it is important to reassess genetic diversity of P. falciparum in Benin to better direct control actions. This study aimed to characterize the allelic polymorphism of msp-1 and msp-2 and determined the multiplicity of infection in P. falciparum isolates collected from children with uncomplicated malaria living in the southern Benin
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