Open Access Research

Pfhrp2 and pfhrp3 polymorphisms in Plasmodium falciparum isolates from Dakar, Senegal: impact on rapid malaria diagnostic tests

Nathalie Wurtz12, Bécaye Fall3, Kim Bui12, Aurélie Pascual124, Mansour Fall5, Cheikhou Camara6, Bakary Diatta5, Khadidiatou Ba Fall7, Pape Saliou Mbaye8, Yaya Diémé3, Raymond Bercion3, Boubacar Wade9, Sébastien Briolant12 and Bruno Pradines1234*

Author Affiliations

1 Unité de Parasitologie, Département d’Infectiologie de Terrain, Institut de Recherche Biomédicale des Armées, Marseille, France

2 Aix Marseille Université, Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes, UM 63, CNRS 7278, IRD 198, Inserm 1095, Marseille, France

3 Laboratoire d’étude de la chimiosensibilité du paludisme, Fédération des laboratoires, Hôpital Principal de Dakar, Dakar, Sénégal

4 Centre National de référence du Paludisme, Marseille, France

5 Service de réanimation médicale, Hôpital Principal de Dakar, Dakar, Sénégal

6 Service des urgences, Hôpital Principal de Dakar, Dakar, Sénégal

7 Service de pathologie infectieuse, Hôpital Principal de Dakar, Dakar, Sénégal

8 Département de médecine interne et spécialités médicales de pathologie tropicale, Hôpital Principal de Dakar, Dakar, Sénégal

9 Chefferie, Hôpital Principal de Dakar, Dakar, Sénégal

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Malaria Journal 2013, 12:34  doi:10.1186/1475-2875-12-34

Published: 24 January 2013

Abstract

Background

An accurate diagnosis is essential for the rapid and appropriate treatment of malaria. The accuracy of the histidine-rich protein 2 (PfHRP2)-based rapid diagnostic test (RDT) Palutop+4® was assessed here. One possible factor contributing to the failure to detect malaria by this test is the diversity of the parasite PfHRP2 antigens.

Methods

PfHRP2 detection with the Palutop+4® RDT was carried out. The pfhrp2 and pfhrp3 genes were amplified and sequenced from 136 isolates of Plasmodium falciparum that were collected in Dakar, Senegal from 2009 to 2011. The DNA sequences were determined and statistical analyses of the variation observed between these two genes were conducted. The potential impact of PfHRP2 and PfHRP3 sequence variation on malaria diagnosis was examined.

Results

Seven P. falciparum isolates (5.9% of the total isolates, regardless of the parasitaemia; 10.7% of the isolates with parasitaemia ≤0.005% or ≤250 parasites/μl) were undetected by the PfHRP2 Palutop+4® RDT. Low parasite density is not sufficient to explain the PfHRP2 detection failure. Three of these seven samples showed pfhrp2 deletion (2.4%). The pfhrp3 gene was deleted in 12.8%. Of the 122 PfHRP2 sequences, 120 unique sequences were identified. Of the 109 PfHRP3 sequences, 64 unique sequences were identified. Using the Baker’s regression model, at least 7.4% of the P. falciparum isolates in Dakar were likely to be undetected by PfHRP2 at a parasite density of ≤250 parasites/μl (slightly lower than the evaluated prevalence of 10.7%). This predictive prevalence increased significantly between 2009 and 2011 (P = 0.0046).

Conclusion

In the present work, 10.7% of the isolates with a parasitaemia ≤0.005% (≤250 parasites/μl) were undetected by the PfHRP2 Palutop+4® RDT (7.4% by the predictive Baker’model). In addition, all of the parasites with pfhrp2 deletion (2.4% of the total samples) and 2.1% of the parasites with parasitaemia >0.005% and presence of pfhrp2 were not detected by PfHRP2 RDT. PfHRP2 is highly polymorphic in Senegal. Efforts should be made to more accurately determine the prevalence of non-sensitive parasites to pfHRP2.

Keywords:
Malaria; Plasmodium falciparum; Diagnosis; Rapid diagnostic test; Plasmodium falciparum histidine-rich protein 2; Anti-malarial