Molecular Identification and Characterization of Plasmodium ovale curtisi in Field Isolates from Symptomatic Children in North-Central Nigeria.

PURPOSE: Plasmodium ovale is not usually the focus of most malaria research or intervention programmes and has lately been termed the neglected human malaria parasites. The parasite exists as two genetically distinct sympatric species namely P. ovale curtisi and P. ovale wallikeri but information on the distribution of P. ovale sub-species is lacking in Nigeria. The objective of this study, therefore, was aimed at characterizing the P. ovale sub-species in isolates from symptomatic individuals in North-central Nigeria. METHODS: Parasites were identified by light microscopy of Giemsa stained thick and thin blood films. Molecular characterization and confirmation of P. ovale sub-species were done by species-specific nested PCR and sequencing of the small subunit ribosomal RNA (SSUrRNA) gene. RESULTS: A total of 412 children were enrolled into this study of which 88.6% (n = 365) were positive for Plasmodium species by nested PCR and P. falciparum was predominant. Of the 365 isolates, 4 (1.1%) had P. ovale infections and of these, 3 (0.8%) were mixed species infections of P. ovale with P. falciparum. DNA sequence analysis confirmed that all the four P. ovale parasites were P. ovale curtisi as their sequences were 99-100% identical to previously published P. ovale curtisi sequences in the GenBank and they cluster with the P. ovale curtisi sequences by phylogeny. CONCLUSION: Our findings demonstrate the occurrence of P. ovale curtisi in the study area. This has implications for public health and malaria elimination programmes, since they also serve as potential risk to travellers from malaria-free regions.

Genetic diversity and complexity of Plasmodium falciparum infections in the microenvironment among siblings of the same household in North-Central Nigeria.

BACKGROUND: Plasmodium falciparum parasites are known to exhibit extensive genetic diversity in areas of high transmission intensity and infected individuals in such communities often harbour several complex mixtures of parasite clones with different genetic characteristics. However, in the micro-environment, the extent of genetic diversity of P. falciparum parasites remain largely unknown. In this study therefore, the complexity of P. falciparum infections in households was investigated among symptomatic siblings, living under the same roof in north-central Nigeria. METHODS: Children were enrolled into the study if they were at least two from a household and presented with symptoms of uncomplicated malaria. Clinical malaria was confirmed by light microscopy of Giemsa-stained thick and thin blood films. Genomic DNA was isolated from blood spots on filter paper. Molecular characterization of P. falciparum isolates was done by allele-specific nested PCR of the highly polymorphic merozoite surface protein-2 (msp-2) gene. RESULTS: Ninety-three children from 43 households were enrolled into this study. A total of 26 different msp-2 alleles were identified from 215 fragments (range: 180-480 bp). Majority of the isolates [65.6% (n = 61)] were polyclonal infections consisting of 2-6 clones and were significantly more common with the FC27 allelic family (p = 0.036). The multiplicity of infection (MOI) per household ranged from 1.0 to 4.5 while the overall MOI in the study population was 2.31. The pattern of distribution of msp-2 allele types among the households fell into two categories: households where both msp-2 allele types (FC27 and 3D7) were present; households where only one msp-2 allele type (FC27 or 3D7) was present. Majority of the households [88.4% (n = 38)], had both msp-2 allele types but they were disproportionately distributed among the children while in a few households [11.6% (n = 5)], all the children were infected with only one type of msp-2 allele. CONCLUSION: These findings showed that P. falciparum isolates exhibit remarkable degree of genetic diversity in the micro-environment and are composed mainly of multiclonal infections, which is an indication of a high ongoing parasite transmission. This suggests that the micro-environment is an important area of focus for malaria control interventions and for evaluating intervention programmes.

Genetic diversity of Plasmodium falciparum isolates from naturally infected children in north-central Nigeria using the merozoite surface protein-2 as molecular marker.

OBJECTIVE: To characterize the genetic diversity of Plasmodium falciparum (P. falciparum) field isolates in children from Lafia, North-central Nigeria, using the highly polymorphic P. falciparum merozoite surface protein 2 (MSP-2) gene as molecular marker. METHODS: Three hundred and twenty children were enrolled into the study between 2005 and 2006. These included 140 children who presented with uncomplicated malaria at the Dalhatu Araf Specialist Hospital, Lafia and another 180 children from the study area with asymptomatic infection. DNA was extracted from blood spot on filter paper and MSP-2 genes were genotyped using allele-specific nested PCR in order to analyze the genetic diversity of parasite isolates. RESULTS: A total of 31 and 34 distinct MSP-2 alleles were identified in the asymptomatic and uncomplicated malaria groups respectively. No difference was found between the multiplicity of infection in the asymptomatic group and that of the uncomplicated malaria group (P>0.05). However, isolates of the FC27 allele type were dominant in the asymptomatic group whereas isolates of the 3D7 allele type were dominant in the uncomplicated malaria group. CONCLUSIONS: This study showed a high genetic diversity of P. falciparum isolates in North-central Nigeria and is comparable to reports from similar areas with high malaria transmission intensity.