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New molecular settings to support in vivo anti-malarial assays

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dc.affiliation.dptoUC3M. Departamento de Bioingenieríaes
dc.affiliation.grupoinvUC3M. Grupo de Investigación: Biomedical Imaging and Instrumentation Groupes
dc.contributor.authorBahamontes Rosa, Noemí
dc.contributor.authorRodríguez Alejandre, Ane
dc.contributor.authorGómez, Vanesa
dc.contributor.authorViera, Sara
dc.contributor.authorGómez Lorenzo, María G.
dc.contributor.authorSanz Alonso, Laura María
dc.contributor.authorMendoza Losana, Alfonso
dc.date.accessioned2023-11-15T14:36:08Z
dc.date.available2023-11-15T14:36:08Z
dc.date.issued2016-03-08
dc.description.abstractBackground Quantitative real-time PCR (qPCR) is now commonly used as a method to confirm diagnosis of malaria and to differentiate recrudescence from re-infection, especially in clinical trials and in reference laboratories where precise quantification is critical. Although anti-malarial drug discovery is based on in vivo murine efficacy models, use of molecular analysis has been limited. The aim of this study was to develop qPCR as a valid methodology to support pre-clinical anti-malarial models by using filter papers to maintain material for qPCR and to compare this with traditional methods. Methods FTA technology (Whatman) is a rapid and safe method for extracting nucleic acids from blood. Peripheral blood samples from mice infected with Plasmodium berghei, P. yoelii, or P. falciparum were kept as frozen samples or as spots on FTA cards. The extracted genetic material from both types of samples was assessed for quantification by qPCR using sets of specific primers specifically designed for Plasmodium 18S rRNA, LDH, and CytB genes. Results The optimal conditions for nucleic acid extraction from FTA cards and qPCR amplification were set up, and were confirmed to be suitable for parasite quantification using DNA as template after storage at room temperature for as long as 26 months in the case of P. berghei samples and 52 months for P. falciparum and P. yoelii. The quality of DNA extracted from the FTA cards for gene sequencing and microsatellite amplification was also assessed. Conclusions This is the first study to report the suitability of FTA cards and qPCR assay to quantify parasite load in samples from in vivo efficacy models to support the drug discovery process.en
dc.identifier.bibliographicCitationBahamontes-Rosa, N., Alejandre, A. R., Gomez, V., Viera, S., Gomez-Lorenzo, M. G., Sanz-Alonso, L. M., & Mendoza-Losana, A. (2016). New molecular settings to support in vivo anti-malarial assays. Malaria Journal, 15 (1).es
dc.identifier.doihttps://doi.org/10.1186/s12936-016-1205-x
dc.identifier.issn1475-2875
dc.identifier.publicationissue147es
dc.identifier.publicationtitleMalaria Journal (Malaria Journal)en
dc.identifier.publicationvolume15es
dc.identifier.urihttps://hdl.handle.net/10016/38871
dc.identifier.uxxiAR/0000030696
dc.language.isoenges
dc.publisherSpringeres
dc.rights© 2016 Bahamontes-Rosa et al.es
dc.rightsAtribución 3.0 España*
dc.rights.accessRightsopen accesses
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/es/*
dc.subject.ecienciaMedicinaes
dc.subject.otherMalariaes
dc.subject.otherPlasmodium falciparumen
dc.subject.otherPlasmodium yoeliies
dc.subject.otherLasmodium bergheien
dc.subject.otherQuantitative real-time PCRen
dc.subject.otherqPCRen
dc.subject.otherFTAen
dc.titleNew molecular settings to support in vivo anti-malarial assaysen
dc.typeresearch article*
dc.type.hasVersionVoR*
dspace.entity.typePublication
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