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Rapid diagnostic tests for molecular surveillance of Plasmodium falciparum malaria -assessment of DNA extraction methods and field applicability

2013Journal article Open access
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dc.contributor.authorMorris, Ulrika
dc.contributor.authorAydin-Schmidt, Berit
dc.contributor.authorShakely, Deler
dc.contributor.authorMartensson, Andreas
dc.contributor.authorJornhagen, Louise
dc.contributor.authorAli, Abdullah S.
dc.contributor.authorMsellem, Mwinyi I.
dc.contributor.authorPetzold, Max
dc.contributor.authorGil, José Pedro
dc.contributor.authorFerreira, Pedro
dc.contributor.authorBjorkman, Anders
dc.date.accessioned2018-12-07T14:53:33Z
dc.date.available2018-12-07T14:53:33Z
dc.date.issued2013
dc.description.abstractBackground: The need for new malaria surveillance tools and strategies is critical, given improved global malaria control and regional elimination efforts. High quality Plasmodium falciparum DNA can reliably be extracted from malaria rapid diagnostic tests (RDTs). Together with highly sensitive molecular assays, wide scale collection of used RDTs may serve as a modern tool for improved malaria case detection and drug resistance surveillance. However, comparative studies of DNA extraction efficiency from RDTs and the field applicability are lacking. The aim of this study was to compare and evaluate different methods of DNA extraction from RDTs and to test the field applicability for the purpose of molecular epidemiological investigations. Methods: DNA was extracted from two RDT devices (Paracheck-PfW and SD Bioline Malaria Pf/Pan (R)), seeded in vitro with 10-fold dilutions of cultured 3D7 P. falciparum parasites diluted in malaria negative whole blood. The level of P. falciparum detection was determined for each extraction method and RDT device with multiple nested-PCR and real-time PCR assays. The field applicability was tested on 855 paired RDT (Paracheck-Pf) and filter paper (Whatman (R) 3MM) blood samples (734 RDT negative and 121 RDT positive samples) collected from febrile patients in Zanzibar 2010. RDT positive samples were genotyped at four key single nucleotide polymorphisms (SNPs) in pfmdr1 and pfcrt as well as for pfmdr1 copy number, all associated with anti-malarial drug resistance. Results: The P. falciparum DNA detection limit varied with RDT device and extraction method. Chelex-100 extraction performed best for all extraction matrixes. There was no statistically significant difference in PCR detection rates in DNA extracted from RDTs and filter paper field samples. Similarly there were no significant differences in the PCR success rates and genotyping outcomes for the respective SNPs in the 121 RDT positive samples. Conclusions: The results support RDTs as a valuable source of parasite DNA and provide evidence for RDT-DNA extraction for improved malaria case detection, molecular drug resistance surveillance, and RDT quality control.
dc.description.sponsorshipACT Consortium through Bill and Melinda Gates Foundation; Swedish International Development Agency (SIDA) [SWE 2009-193]; Swedish Civil Contingencies Agency (MSB) [2010-7991]; Swedish Medical Research Council (VR) [2009-3785]; Goljes Foundation
dc.description.versioninfo:eu-repo/semantics/publishedVersion
dc.identifier.doi10.1186/1475-2875-12-106
dc.identifier.issn1475-2875
dc.identifier.urihttp://hdl.handle.net/10400.1/11564
dc.language.isoeng
dc.peerreviewedyes
dc.publisherBMC
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectReal-Time Pcr
dc.subjectMixed infections
dc.subjectBlood spots
dc.subjectPrevalence
dc.subjectPfmdr1
dc.subjectAmodiaquine
dc.subjectResistance
dc.subjectMutations
dc.subjectMarkers
dc.subjectAssay
dc.titleRapid diagnostic tests for molecular surveillance of Plasmodium falciparum malaria -assessment of DNA extraction methods and field applicability
dc.typejournal article
dspace.entity.typePublication
oaire.citation.startPage106
oaire.citation.titleMalaria Journal
oaire.citation.volume12
person.familyNameGil
person.givenNameJosé Pedro
person.identifier.ciencia-idD01A-B30E-BCD5
person.identifier.orcid0000-0002-6107-9379
person.identifier.scopus-author-id7201625436
rcaap.rightsopenAccess
rcaap.typearticle
relation.isAuthorOfPublicationcb728715-0e4c-4ae5-9e21-b6a8f35a8313
relation.isAuthorOfPublication.latestForDiscoverycb728715-0e4c-4ae5-9e21-b6a8f35a8313

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