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Real-time PCR/MCA assay using fluorescence resonance energy transfer for the genotyping of resistance related DHPS-540 mutations in Plasmodium falciparum

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dc.contributor.author Mens, P. F. en_US
dc.contributor.author Van Overmeir, C. en_US
dc.contributor.author Bonnet, M. en_US
dc.contributor.author Dujardin, J. C. en_US
dc.contributor.author D'Alessandro, U. en_US
dc.date.accessioned 2008-04-08T10:29:44Z
dc.date.available 2008-04-08T10:29:44Z
dc.date.issued 2008
dc.identifier.issn 1475-2875
dc.identifier.doi http://dx.doi.org/10.1186/1475-287-7-48
dc.identifier.other ITG-P1B en_US
dc.identifier.other ITG-P2B en_US
dc.identifier.other ITG-P4A en_US
dc.identifier.other ITG-PLA en_US
dc.identifier.other PARAS en_US
dc.identifier.other U-MALAR en_US
dc.identifier.other U-PROTO en_US
dc.identifier.other JIF en_US
dc.identifier.other DOI en_US
dc.identifier.other UPD1 en_US
dc.identifier.other FTA en_US
dc.identifier.other ELECTRONIC en_US
dc.identifier.other ABSTRACT en_US
dc.identifier.uri http://hdl.handle.net/10390/2229
dc.description.abstract BACKGROUND: Sulphadoxine-pyrimethamine has been abandoned as first- or second-line treatment by most African malaria endemic countries in favour of artemisinin-based combination treatments, but the drug is still used as intermittent preventive treatment during pregnancy. However, resistance to sulphadoxine-pyrimethamine has been increasing in the past few years and, although the link between molecular markers and treatment failure has not been firmly established, at least for pregnant women, it is important to monitor such markers. METHODS: This paper reports a novel sensitive, semi-quantitative and specific real-time PCR and melting curve analysis (MCA) assay using fluorescence resonance energy transfer (FRET) for the detection of DHPS-540, an important predictor for SP resistance. FRET/MCA was evaluated using 78 clinical samples from malaria patients and compared to PCR-RFLP. RESULTS: Sixty-two samples were in perfect agreement between both assays. One sample showed a small wild type signal with FRET/MCA that indicates a polyclonal infection. Four samples were not able to generate enough material in both assays to distinguish mutant from wild-type infection, six samples gave no signal in PCR-RFLP and five samples gave no amplification in FRET/MCA. CONCLUSION: FRET/MCA is an effective tool for the identification of SNPs in drug studies and epidemiological surveys on resistance markers in general and DHPS-540 mutation in particular. en_US
dc.language English en_US
dc.publisher BioMed Central en_US
dc.subject Protozoal diseases en_US
dc.subject Malaria en_US
dc.subject Plasmodium falciparum en_US
dc.subject Drug resistance en_US
dc.subject Mutations en_US
dc.subject DHPS-540 en_US
dc.subject Genotyping en_US
dc.subject Real-time en_US
dc.subject Polymerase chain reaction en_US
dc.subject PCR en_US
dc.subject Melting curve analysis en_US
dc.subject MCA en_US
dc.title Real-time PCR/MCA assay using fluorescence resonance energy transfer for the genotyping of resistance related DHPS-540 mutations in Plasmodium falciparum en_US
dc.type Article-E en_US
dc.citation.issue 48 en_US
dc.citation.jtitle Malaria Journal en_US
dc.citation.volume 7 en_US
dc.citation.pages 7 en_US
dc.publisher.place London en_US
dc.identifier.pmid http://www.ncbi.nlm.nih.gov/pubmed/18346279
dc.identifier.url http://www.malariajournal.com/content/7/1/48
dc.citation.jabbreviation Malaria J en_US


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