High frequency of the Duffy-negative genotype and absence of Plasmodium vivax infections in Ghana

Abstract

Background: Recent studies from different malaria-endemic regions including western Africa have now shown that Plasmodium vivax can infect red blood cells (RBCs) and cause clinical disease in Duffy-negative people, though the Duffy-negative phenotype was thought to confer complete refractoriness against blood invasion with P. vivax. The actual prevalence of P. vivax in local populations in Ghana is unknown and little information is available about the distribution of Duffy genotypes. The aim of this study was to assess the prevalence of P. vivax in both asymptomatic and symptomatic outpatients and the distribution of Duffy genotypes in Ghana.

Methods: DNA was extracted from dried blood spots (DBS) collected from 952 subjects (845 malaria patients and 107 asymptomatic persons) from nine locations in Ghana. Plasmodium species identification was carried out by nested polymerase chain reaction (PCR) amplification of the small-subunit (SSU) rRNA genes. For P. vivax detection, a second PCR of the central region of the Pvcsp gene was carried out. Duffy blood group genotyping was performed by allele-specific PCR to detect the presence of the FY^ES allele.

Results: No cases of P. vivax were detected in any of the samples by both PCR methods used. Majority of infections (542, 94.8%) in the malaria patient samples were due to P. falciparum with only 1 infection (0.0017%) due to Plasmodium malariae, and 2 infections (0.0034%) due to Plasmodium ovale. No case of mixed infection was identified. Of the samples tested for the FY^ES allele from all the sites, 90.5% (862/952) had the FY^ES allele. All positive samples were genotyped as FY*B-33/FY*B-33 (Duffy-negative homozygous) and therefore classified as Fy(a-b-).

Conclusions: No cases of P. vivax were detected by both PCRs and majority of the subjects tested carried the FY^ES allele. The lack of P. vivax infections observed can be attributed to the high frequency of the FY^ES allele that silences erythroid expression of the Duffy. These results provide insights on the host susceptibility for P. vivax infections that had not been investigated in Ghana before.

Keywords: Duffy blood group; Duffy-negative; Ghana; Malaria; Plasmodium vivax.

Fig. 1

A map of Ghana showing the 9 collection sites. U urban, R rural (Adapted from Duah et al. [28])

Fig. 2

Example of an ethidium bromide-stained 1.0% agarose gel electrophoresis of allele specific PCR products for Duffy genotyping. Lanes 1 and 2 show PCR positive for both internal control and Duffy allele; Lane 3 shows a negative sample; Lane 4 shows PCR positive for only Duffy allele; Lanes 5 and 6 show PCR positive for only internal control. M = 100 bp ladder (NEB)