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

Clark, D.; Zhu, D.; Calderón Sánchez, Maritza Mercedes; Sterling, C.; Gilman, Robert Hugh; Traianou, A.; Holtz, M.; Acosta, J.; Málaga, E.; Ramal, C.; Fortuny, E.; Diestra, A.; Valencia, G.; Gandarilla, O.; Bowman, N.; Steinberg, H.; Reimer-McAtee, M.; Angulo, N.; Bok, J.; Choi, J.; Schiaffino Salazar, Francesca; Ticona, E.; Jiménez, J.; Calla Choque, Jaeson Santos; Ferradas, C.; Pinedo, L.C.; Mayta, H.; Clark, T.; Gutierrez Loli, Renzo Marcelo; Galdos, G.; Trompeter, G.; Bern, C.; Colanzi, R.; Dorn, M.; Sanchez, L.

dc.contributor.author	Gutierrez Loli, Renzo Marcelo
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	Calderón Sánchez, Maritza Mercedes
dc.contributor.author	Calla Choque, Jaeson Santos
dc.contributor.author	Sterling, C.
dc.contributor.author	Gilman, Robert Hugh
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 Salazar, Francesca
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.ispartofseries	American Journal of Tropical Medicine and Hygiene (ASTMH Journal)
dc.rights	info:eu-repo/semantics/restrictedAccess
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
dc.relation.issn	1476-1645