Plasmodium falciparum Allelic Diversity: A Comparison of DNA Extraction from Isolates Collected on Rapid Diagnostic Tests (Rdts) and Filter Paper

Denise Patricia  Mawili-Mboumba; Marie Louise  Tshibola Mbuyi; Noe Patrick  M’bondoukwe; Marielle Karine  Bouyou-Akotet

doi:10.18502/ijpa.v16i4.7867

Denise Patricia Mawili-Mboumba Department of Parasitology-Mycology, Faculty of Medicine, Université des Sciences de la Santé, BP4009, Libreville, Gabon
Marie Louise Tshibola Mbuyi Department of Parasitology-Mycology, Faculty of Medicine, Université des Sciences de la Santé, BP4009, Libreville, Gabon
Noe Patrick M’bondoukwe Department of Parasitology-Mycology, Faculty of Medicine, Université des Sciences de la Santé, BP4009, Libreville, Gabon
Marielle Karine Bouyou-Akotet Department of Parasitology-Mycology, Faculty of Medicine, Université des Sciences de la Santé, BP4009, Libreville, Gabon

DOI: https://doi.org/10.18502/ijpa.v16i4.7867

Keywords: DNA; Genetic diversity; Rapid diagnostic tests; Malaria

Abstract

Background: To perform molecular epidemiologic studies based on large cohorts, material such as RDTs or filter papers are useful for biological sample collection and extraction of RNA or DNA of good quality. Thus, we aimed to assess the quality of DNA extracted from malaria rapid diagnostic tests (RDTs) stored at various temperatures for the analysis of Plasmodium falciparum genetic diversity.

Methods: Febrile patients benefitted from free malaria diagnosis using microscopy in a malaria sentinel site, at the Regional Hospital Estuaire-Melen, in Gabon, in 2015. P. falciparum isolates were collected onto one filter paper and 2 similar RDTs devices (Acon®) per patient. Nucleic acids were extracted with QiAmp Qiagen kit from paper and RDTs and the quality of the DNA was analyzed by msp1 gene amplification.

Results: Msp1gene amplification was achieved in nucleic acids extracted from all filter papers and RDTs devices (n = 45, 100%). K1 alleles were detected in 93.3% (n = 42/45) of the samples and Mad20 alleles in 73.3% (n = 33/45). The number and the intensity of K1 and/or Mad20 fragments were comparable according to the sample collection material and the storage conditions (room temperature vs -20°C) of the samples. The size of the fragments indicating allelic diversity was comparable in 80% (n=36) of the samples.

Conclusion: These data show that RDTs are a valuable source of DNA for malaria parasite genetic polymorphism analysis. Storage conditions of the devices did not influence the quality of DNA extracted from RDTs device, although some alleles may be missed.