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Analytical Validation of Quantitative Real-Time PCR Methods for Quantification of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients

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dc.contributor.author Ramirez, Juan Carlos
dc.contributor.author Cura, Carolina Ines
dc.contributor.author da Cruz Moreira, Otacilio
dc.contributor.author Lages-Silva, Eliane
dc.contributor.author Juiz, Natalia
dc.contributor.author Velazquez, Elsa
dc.contributor.author Ramirez, Juan David
dc.contributor.author Alberti, Anahi
dc.contributor.author Pavia, Paula
dc.contributor.author Flores-Chavez, Maria Delmans
dc.contributor.author Munoz-Calderon, Arturo
dc.contributor.author Perez-Morales, Deyanira
dc.contributor.author Santalla, Jose
dc.contributor.author Marcos da Matta Guedes, Paulo
dc.contributor.author Peneau, Julie
dc.contributor.author Marcet, Paula
dc.contributor.author Padilla, Carlos
dc.contributor.author Cruz-Robles, David
dc.contributor.author Valencia, Edward
dc.contributor.author Crisante, Gladys Elena
dc.contributor.author Greif, Gonzalo
dc.contributor.author Zulantay, Ines
dc.contributor.author Costales, Jaime Alfredo
dc.contributor.author Alvarez-Martinez, Miriam
dc.contributor.author Martinez, Norma Edith
dc.contributor.author Villarroel, Rodrigo
dc.contributor.author Villarroel, Sandro
dc.contributor.author Sanchez, Zunilda
dc.contributor.author Bisio, Margarita
dc.contributor.author Parrado, Rudy
dc.contributor.author Maria da Cunha Galvao, Lucia
dc.contributor.author Jacome da Camara, Antonia Claudia
dc.contributor.author Espinoza, Bertha
dc.contributor.author Alarcon de Noya, Belkisyole
dc.contributor.author Puerta, Concepcion
dc.contributor.author Riarte, Adelina
dc.contributor.author Diosque, Patricio
dc.contributor.author Sosa-Estani, Sergio
dc.contributor.author Guhl, Felipe
dc.contributor.author Ribeiro, Isabela
dc.contributor.author Aznar, Christine
dc.contributor.author Britto, Constanca
dc.contributor.author Yadon, Zaida Estela
dc.contributor.author Schijman, Alejandro G.
dc.date.accessioned 2019-02-06T14:57:41Z
dc.date.available 2019-02-06T14:57:41Z
dc.date.issued 2015
dc.identifier.uri https://hdl.handle.net/20.500.12866/5491
dc.description.abstract An international study was performed by 26 experienced PCR laboratories from 14 countries to assess the performance of duplex quantitative real-time PCR (qPCR) strategies on the basis of TaqMan probes for detection and quantification of parasitic loads in peripheral blood samples from Chagas disease patients. Two methods were studied: Satellite DNA (SatDNA) qPCR and kinetoplastid DNA (kDNA) qPCR. Both methods included an internal amplification control. Reportable range, analytical sensitivity, limits of detection and quantification, and precision were estimated according to international guidelines. In addition, inclusivity and exclusivity were estimated with DNA from stocks representing the different Trypanosoma cruzi discrete typing units and Trypanosoma rangeli and Leishmania spp. Both methods were challenged against 156 blood samples provided by the participant laboratories, including samples from acute and chronic patients with varied clinical findings, infected by oral route or vectorial transmission. kDNA qPCR showed better analytical sensitivity than SatDNA qPCR with limits of detection of 0.23 and 0.70 parasite equivalents/mL, respectively. Analyses of clinical samples revealed a high concordance in terms of sensitivity and parasitic loads determined by both SatDNA and kDNA qPCRs. This effort is a major step toward international validation of qPCR methods for the quantification of T. cruzi DNA in human blood samples, aiming to provide an accurate surrogate biomarker for diagnosis and treatment monitoring for patients with Chagas disease. en_US
dc.language.iso eng
dc.publisher Elsevier
dc.relation.ispartof urn:issn:1943-7811
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Humans en_US
dc.subject Sensitivity and Specificity en_US
dc.subject International Cooperation en_US
dc.subject Real-Time Polymerase Chain Reaction/methods en_US
dc.subject Molecular Typing en_US
dc.subject Chagas Disease/blood/diagnosis/genetics/parasitology en_US
dc.subject DNA, Protozoan/analysis/isolation & purification en_US
dc.subject Laboratory Proficiency Testing en_US
dc.subject Parasitemia/blood/diagnosis/genetics en_US
dc.subject Trypanosoma cruzi/genetics/isolation & purification en_US
dc.title Analytical Validation of Quantitative Real-Time PCR Methods for Quantification of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1016/j.jmoldx.2015.04.010
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.02.00 es_PE

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