Asymptomatic malaria infections and Pfmdr1 mutations in an endemic area of Nigeria

Abstract

Background: Malaria eradication globally is yet to be achieved and transmission is sustained in many endemic countries. Plasmodium falciparum continues to develop resistance to currently available anti-malarial drugs, posing great problems for malaria elimination. This study evaluates the frequencies of asymptomatic infection and multidrug resistance-1 (mdr-1) gene mutations in parasite isolates, which form the basis for understanding persistently high incidence in South West, Nigeria.

Methods: A total of 535 individuals aged from 6 months were screened during the epidemiological survey evaluating asymptomatic transmission. Parasite prevalence was determined by histidine-rich protein II rapid detection kit (RDT) in healthy individuals. Plasmodium falciparum mdr-1 gene mutations were detected by polymerase chain reaction (PCR) followed by restriction enzyme digest and electrophoresis to determine polymorphism in parasite isolates. Sequencing was done to confirm polymorphism. Proportions were compared using Chi-square test at p value < 0.05.

Results: Malaria parasites were detected by RDT in 204 (38.1%) individuals. Asymptomatic infection was detected in 117 (57.3%) and symptomatic malaria confirmed in 87 individuals (42.6%). Overall, individuals with detectable malaria by RDT was significantly higher in individuals with symptoms, 87 of 197 (44.2%), than asymptomatic persons; 117 of 338 (34.6%), p = 0.02. In a sub-set of 75 isolates, 18(24%) and 14 (18.6%) individuals had Pfmdr1 86Y and 1246Y mutations.

Conclusions: There is still high malaria transmission rate in Nigeria with higher incidence of asymptomatic infections. These parasites harbour mutations on Pfmdr1 which contribute to artemisinin partner drug resistance; surveillance strategies to reduce the spread of drug resistance in endemic areas are needed to eliminate the reservoir of malaria parasites that can mitigate the eradication of malaria in Nigeria.

Keywords: Asymptomatic malaria; Drug resistance; Nigeria; Pfmdr1; Plasmodium falciparum.

Fig. 1

Flow chart for the study

Fig. 2

Prevalence of mutant alleles detected at codon 86 (24%) and 1246 (19%) of Pfmdr 1 gene in parasite isolates during the study

Fig. 3

Restriction digest product for Pfmdr1 SNP detection at codon 86 (a) and 1246 (b)

Fig. 4

Screenshot of alignment of Pfmdr1 gene flanking codon 86 in 2 isolates (with mutant allele- tyrosine) compared to reference 3D7 strain (sensitive allele-asparagine)