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RDTs as a source of DNA to study Plasmodium falciparum drug resistance in isolates from Senegal and the Comoros Islands
BACKGROUND: The World Health Organization has recommended rapid diagnostic tests (RDTs) for use in the diagnosis of suspected malaria cases. In addition to providing quick and accurate detection of Plasmodium parasite proteins in the blood, these tests can be used as sources of DNA for further genet...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4587814/ https://www.ncbi.nlm.nih.gov/pubmed/26415927 http://dx.doi.org/10.1186/s12936-015-0861-6 |
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author | Papa Mze, Nasserdine Ndiaye, Yaye Die Diedhiou, Cyrille K. Rahamatou, Silai Dieye, Baba Daniels, Rachel F. Hamilton, Elizabeth J. Diallo, Mouhamadou Bei, Amy K. Wirth, Dyann F. Mboup, Souleymane Volkman, Sarah K. Ahouidi, Ambroise D. Ndiaye, Daouda |
author_facet | Papa Mze, Nasserdine Ndiaye, Yaye Die Diedhiou, Cyrille K. Rahamatou, Silai Dieye, Baba Daniels, Rachel F. Hamilton, Elizabeth J. Diallo, Mouhamadou Bei, Amy K. Wirth, Dyann F. Mboup, Souleymane Volkman, Sarah K. Ahouidi, Ambroise D. Ndiaye, Daouda |
author_sort | Papa Mze, Nasserdine |
collection | PubMed |
description | BACKGROUND: The World Health Organization has recommended rapid diagnostic tests (RDTs) for use in the diagnosis of suspected malaria cases. In addition to providing quick and accurate detection of Plasmodium parasite proteins in the blood, these tests can be used as sources of DNA for further genetic studies. As sulfadoxine-pyrimethamine is used currently for intermittent presumptive treatment of pregnant women in both Senegal and in the Comoros Islands, resistance mutations in the dhfr and dhps genes were investigated using DNA extracted from RDTs. METHODS: The proximal portion of the nitrocellulose membrane of discarded RDTs was used for DNA extraction. This genomic DNA was amplified using HRM to genotype the molecular markers involved in resistance to sulfadoxine-pyrimethamine: dhfr (51, 59, 108, and 164) and dhps (436, 437, 540, 581, and 613). Additionally, the msp1 and msp2 genes were amplified to determine the average clonality between Grande-Comore (Comoros) and Thiès (Senegal). RESULTS: A total of 201 samples were successfully genotyped at all codons by HRM; whereas, in 200 msp1 and msp2 genes were successfully amplified and genotyped by nested PCR. A high prevalence of resistance mutations were observed in the dhfr gene at codons 51, 59, and 108 as well as in the dhps gene at codons 437 and 436. A novel mutant in dhps at codon positions 436Y/437A was observed. The dhfr I164L codon and dhps K540 and dhps A581G codons had 100 % wild type alleles in all samples. CONCLUSION: The utility of field-collected RDTs was validated as a source of DNA for genetic studies interrogating frequencies of drug resistance mutations, using two different molecular methods (PCR and High Resolution Melting). RDTs should not be discarded after use as they can be a valuable source of DNA for genetic and epidemiological studies in sites where filter paper or venous blood collected samples are nonexistent. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12936-015-0861-6) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4587814 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-45878142015-09-30 RDTs as a source of DNA to study Plasmodium falciparum drug resistance in isolates from Senegal and the Comoros Islands Papa Mze, Nasserdine Ndiaye, Yaye Die Diedhiou, Cyrille K. Rahamatou, Silai Dieye, Baba Daniels, Rachel F. Hamilton, Elizabeth J. Diallo, Mouhamadou Bei, Amy K. Wirth, Dyann F. Mboup, Souleymane Volkman, Sarah K. Ahouidi, Ambroise D. Ndiaye, Daouda Malar J Research BACKGROUND: The World Health Organization has recommended rapid diagnostic tests (RDTs) for use in the diagnosis of suspected malaria cases. In addition to providing quick and accurate detection of Plasmodium parasite proteins in the blood, these tests can be used as sources of DNA for further genetic studies. As sulfadoxine-pyrimethamine is used currently for intermittent presumptive treatment of pregnant women in both Senegal and in the Comoros Islands, resistance mutations in the dhfr and dhps genes were investigated using DNA extracted from RDTs. METHODS: The proximal portion of the nitrocellulose membrane of discarded RDTs was used for DNA extraction. This genomic DNA was amplified using HRM to genotype the molecular markers involved in resistance to sulfadoxine-pyrimethamine: dhfr (51, 59, 108, and 164) and dhps (436, 437, 540, 581, and 613). Additionally, the msp1 and msp2 genes were amplified to determine the average clonality between Grande-Comore (Comoros) and Thiès (Senegal). RESULTS: A total of 201 samples were successfully genotyped at all codons by HRM; whereas, in 200 msp1 and msp2 genes were successfully amplified and genotyped by nested PCR. A high prevalence of resistance mutations were observed in the dhfr gene at codons 51, 59, and 108 as well as in the dhps gene at codons 437 and 436. A novel mutant in dhps at codon positions 436Y/437A was observed. The dhfr I164L codon and dhps K540 and dhps A581G codons had 100 % wild type alleles in all samples. CONCLUSION: The utility of field-collected RDTs was validated as a source of DNA for genetic studies interrogating frequencies of drug resistance mutations, using two different molecular methods (PCR and High Resolution Melting). RDTs should not be discarded after use as they can be a valuable source of DNA for genetic and epidemiological studies in sites where filter paper or venous blood collected samples are nonexistent. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12936-015-0861-6) contains supplementary material, which is available to authorized users. BioMed Central 2015-09-29 /pmc/articles/PMC4587814/ /pubmed/26415927 http://dx.doi.org/10.1186/s12936-015-0861-6 Text en © Papa Mze et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Papa Mze, Nasserdine Ndiaye, Yaye Die Diedhiou, Cyrille K. Rahamatou, Silai Dieye, Baba Daniels, Rachel F. Hamilton, Elizabeth J. Diallo, Mouhamadou Bei, Amy K. Wirth, Dyann F. Mboup, Souleymane Volkman, Sarah K. Ahouidi, Ambroise D. Ndiaye, Daouda RDTs as a source of DNA to study Plasmodium falciparum drug resistance in isolates from Senegal and the Comoros Islands |
title | RDTs as a source of DNA to study Plasmodium falciparum drug resistance in isolates from Senegal and the Comoros Islands |
title_full | RDTs as a source of DNA to study Plasmodium falciparum drug resistance in isolates from Senegal and the Comoros Islands |
title_fullStr | RDTs as a source of DNA to study Plasmodium falciparum drug resistance in isolates from Senegal and the Comoros Islands |
title_full_unstemmed | RDTs as a source of DNA to study Plasmodium falciparum drug resistance in isolates from Senegal and the Comoros Islands |
title_short | RDTs as a source of DNA to study Plasmodium falciparum drug resistance in isolates from Senegal and the Comoros Islands |
title_sort | rdts as a source of dna to study plasmodium falciparum drug resistance in isolates from senegal and the comoros islands |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4587814/ https://www.ncbi.nlm.nih.gov/pubmed/26415927 http://dx.doi.org/10.1186/s12936-015-0861-6 |
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