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Development of a novel protocol based on blood clot to improve the sensitivity of qPCR detection of toxoplasma gondii in peripheral blood specimens

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dc.contributor.author Gutierrez-Loli, R.
dc.contributor.author Ferradas, C.
dc.contributor.author Diestra, A.
dc.contributor.author Traianou, A.
dc.contributor.author Bowman, N.
dc.contributor.author Bok, J.
dc.contributor.author Reimer-McAtee, M.
dc.contributor.author Ramal, C.
dc.contributor.author Ticona, E.
dc.contributor.author Steinberg, H.
dc.contributor.author Mayta, H.
dc.contributor.author Calderon, M.
dc.contributor.author Calla-Choque, J.S.
dc.contributor.author Sterling, C.
dc.contributor.author Gilman, R.H.
dc.contributor.author Pinedo, L.C.
dc.contributor.author Valencia, G.
dc.contributor.author Sanchez, L.
dc.contributor.author Málaga, E.
dc.contributor.author Zhu, D.
dc.contributor.author Jiménez, J.
dc.contributor.author Bern, C.
dc.contributor.author Angulo, N.
dc.contributor.author Schiaffino, F.
dc.contributor.author Acosta, J.
dc.contributor.author Holtz, M.
dc.contributor.author Clark, D.
dc.contributor.author Clark, T.
dc.contributor.author Trompeter, G.
dc.contributor.author Choi, J.
dc.contributor.author Gandarilla, O.
dc.contributor.author Dorn, M.
dc.contributor.author Fortuny, E.
dc.contributor.author Galdos, G.
dc.contributor.author Colanzi, R.
dc.date.accessioned 2019-12-06T20:57:49Z
dc.date.available 2019-12-06T20:57:49Z
dc.date.issued 2019
dc.identifier.uri https://hdl.handle.net/20.500.12866/7434
dc.description.abstract Quantitative polymerase chain reaction (qPCR) for Toxoplasma gondii multicopy genes has emerged as a promising strategy for sensitive detection of parasite DNA. qPCR can be performed from blood samples, which are minimally invasive to collect. However, there is no consensus about what type of blood specimen yields the best sensitivity. The development of a novel protocol for qPCR detection of T. gondii using blood clot, involving an appropriate DNA extraction method and the use of an internal amplification control to monitor the reaction is presented in the current study. Assays directed to the B1 and REP529 genes were performed in spiked specimens of whole blood, guanidine–ethylenediaminetetraacetic acid blood, and clot. The clot-based qPCR was shown to be more sensitive when compared with other types of specimens, detecting five and 0.05 T. gondii genomes, using B1 and REP529 targets, respectively. Finally, a comparative analysis with samples from HIV patients with clinical suspicion of toxoplasmosis was performed, demonstrating the detection of four positive suspected cases with clots compared with only one using guanidine–ethylenediaminetetraacetic acid blood. The high analytical sensitivity and the cost-effective advantages offered by clot supports this methodology as a good laboratory tool to monitor parasite burden. en_US
dc.language.iso eng
dc.publisher American Society of Tropical Medicine and Hygiene
dc.relation.ispartof urn:issn:1476-1645
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject adult en_US
dc.subject Adult en_US
dc.subject animal cell en_US
dc.subject Article en_US
dc.subject B1 gene en_US
dc.subject blood en_US
dc.subject blood clot en_US
dc.subject blood sampling en_US
dc.subject clinical article en_US
dc.subject control strategy en_US
dc.subject controlled study en_US
dc.subject DNA extraction en_US
dc.subject DNA, Protozoan en_US
dc.subject edetic acid en_US
dc.subject gene en_US
dc.subject genetics en_US
dc.subject genome en_US
dc.subject Genome, Protozoan en_US
dc.subject guanidine en_US
dc.subject HIV Infections en_US
dc.subject human en_US
dc.subject Human immunodeficiency virus infected patient en_US
dc.subject Human immunodeficiency virus infection en_US
dc.subject Humans en_US
dc.subject internal amplification control en_US
dc.subject isolation and purification en_US
dc.subject molecular diagnosis en_US
dc.subject Molecular Diagnostic Techniques en_US
dc.subject nonhuman en_US
dc.subject parasite load en_US
dc.subject Parasite Load en_US
dc.subject parasitology en_US
dc.subject polymerase chain reaction en_US
dc.subject Polymerase Chain Reaction en_US
dc.subject procedures en_US
dc.subject protozoal DNA en_US
dc.subject quantitative analysis en_US
dc.subject REP529 gene en_US
dc.subject sensitivity and specificity en_US
dc.subject Sensitivity and Specificity en_US
dc.subject thrombosis en_US
dc.subject Thrombosis en_US
dc.subject Toxoplasma en_US
dc.subject Toxoplasma gondii en_US
dc.subject toxoplasmosis en_US
dc.subject Toxoplasmosis en_US
dc.subject young adult en_US
dc.subject Young Adult en_US
dc.title Development of a novel protocol based on blood clot to improve the sensitivity of qPCR detection of toxoplasma gondii in peripheral blood specimens en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.4269/ajtmh.17-0920
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.03.06 es_PE
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.03.06


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