Original Article

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


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.

1. Cnops L, Van Esbroeck M, Bottieau E, Jacobs J. Giemsa-stained thick blood films as a source of DNA for Plasmodium spe-cies-specific real-time PCR. Malar J. 2010; 9:370-376.
2. Cnops L, Jacobs J, Van Esbroeck M. Val-idation of a four-primer real-time PCR as a diagnostic tool for single and mixed Plas-modium infections. Clin Microbiol Infect. 2011; 17: 1101-7.
3. Bereczky S, Mårtensson A, Pedro Gil J, Farnert A. Rapid DNA extraction from ar-chive blood spots on filter paper for genotyping of Plasmodium falciparum. Am J Trop Med Hyg. 2005; 72: 249–251.
4. Zainabadi, K, Adams M, Han ZY. Lwin HW, Han KT, Ouattara A, Thura S, Plowe CV, Nyunt MM. A novel method for ex-tracting nucleic acids from dried blood spots for ultrasensitive detection of low-density Plasmodium falciparum and Plasmodium vivax infections. Malar J. 2017; 16, 377.
5. 5- Wongsrichanalai C, Barcus MJ, Muth S, Sutamihardja A, Wernsdorfer WH. A re-view of malaria diagnostic tools: micros-copy and rapid diagnostic test (RDT). Am J Trop Med Hyg 2007; 77:119-127.
6. Cnops L, Boderie, M, Gillet P, Van Esbroeck M, Jacobs J. Rapid diagnostic tests as a source of DNA for Plasmodium species-specific real-time PCR. Malar J. 2011;10:67-77.
7. Ishengoma DS, Lwitiho S, Madebe RA, et al. Using rapid diagnostic tests as source of malaria parasite DNA for molecular analyses in the era of declining malaria prevalence. Malar J. 2011; 10:6-12.
8. Morris U, Aydin-Schmidt B, Shakely D, Martensson A, Jornhagen L, Ali AS. Rapid diagnostic tests for molecular surveillance of Plasmodium falciparum malaria-assessment of DNA extraction methods and field ap-plicability. Malar J. 2013; 12:106-111.
9. Papa Mze N, Ndiaye YD, Diedhiou CK, et al. RDTs as a source of DNA to study Plasmodium falciparum drug resistance in iso-lates from Senegal and the Comoros Is-lands. 2015; Malar J 4:373-380.
10. Planche T, Krishna S, Kombila M, Engel K, Faucher JF, Ngou-Milama E, Kremsner PG. Comparison of methods for the rap-id laboratory assessment of children with malaria. Am J Trop Med Hyg 2001; 65: 599-602.
11. Mawili-Mboumba DP, Bouyou Akotet MK, Ngoungou EB, Kombila M. Evalua-tion of rapid diagnostic tests for malaria case management in Gabon. Diagn Mi-crobiol Infect Dis 2010; 66: 162-168.
12. Mawili-Mboumba DP, Mbondoukwe N, Adande E, Bouyou-Akotet MK. Allelic Diversity of MSP1 Gene in Plasmodium falci-parum from Rural and Urban Areas of Gabon. Korean J Parasitol 2015; 53: 413-419.
13. Bouyou-Akotet MK, M’Bondoukwé NP, Mawili-Mboumba DP. Genetic polymor-phism of merozoite surface protein-1 in Plasmodium falciparum isolates from patients with mild to severe malaria in Libreville, Gabon. Parasite 2015; 22: 12-20.
14. Aubouy A, Migot-Nabias F, Deloron P. Polymorphism in two merozoite surface proteins of Plasmodium falciparum isolates from Gabon. Malar J 2003; 2: 12-17.
15. Ndong Ngomo JM, M’Bondoukwe NP, Yavo W, Bongho Mavoungou LC, et al. Spatial and temporal distribution of Pfmsp1 and Pfmsp2 alleles and genetic profile change of Plasmodium falciparum populations in Gabon. 2018; Acta Tropica 178: 27–33.
16. Mawili-Mboumba DP, Bouyou-Akotet MK, Offouga Mbouoronde C, Kombila M. Analysis of malaria diagnosis discrep-ancies between RDTs and microscopy by nested PCR. J. Biomed Sci Eng. 2013; 6: 967-972.
17. Guirou EA, Schindler T, Hosch S, et al. Molecular malaria surveillance using a nov-el protocol for extraction and analysis of nucleic acids retained on used rapid diag-nostic tests. Sci Rep. 2020; 10 (1):12305.
18. Nguyen TT, Nzigou Mombo B, Lalremru-ata A, et al. DNA recovery from ar-chived RDTs for genetic characterization of Plasmodium falciparum in a routine setting in Lambarene, Gabon. Malar J. 2019;18 (1):336.
IssueVol 16 No 4 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v16i4.7867
DNA Genetic diversity Rapid diagnostic tests Malaria

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Mawili-Mboumba DP, Tshibola Mbuyi ML, M’Bondoukwe NP, Bouyou Akotet MK. Plasmodium falciparum Allelic Diversity: A Comparison of DNA Extraction from Isolates Collected on Rapid Diagnostic Tests (Rdts) and Filter Paper. Iran J Parasitol. 2021;16(4):555-559.