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

dc.creatorRamirez, Juan Carlos
dc.creatorCura, Carolina Inés
dc.creatorMoreira, Otacilio da Cruz
dc.creatorSilva, Eliane Lages
dc.creatorJuiz, Natalia
dc.creatorVelásquez, Elsa
dc.creatorRamirez, Juan David
dc.creatorAlberti, Anahi
dc.creatorPavia, Paula
dc.creatorFlores-Chávez, Maria Delmans
dc.creatorMuñoz-Calderón, Arturo
dc.creatorPérez-Morales, Deyanira
dc.creatorSantalia, José
dc.creatorGuedes, Paulo Marcos da Matta
dc.creatorPeneau, Julie
dc.creatorMarcet, Paula
dc.creatorPadilla, Carlos
dc.creatorCruz-Robles, David
dc.creatorValencia, Edward
dc.creatorCrisante, Gladys Elena
dc.creatorGreif, Gonzalo
dc.creatorZulantay, Inés
dc.creatorCostales, Jaime Alfredo
dc.creatorAlvarez-Martínez, Miriam
dc.creatorMartinez, Norma Edith
dc.creatorVillarroel, Rodrigo
dc.creatorVillarroel, Sandro
dc.creatorSánchez, Zunilda
dc.creatorBisio, Margarita
dc.creatorParrado, Rudy
dc.creatorGalvão, Lúcia Maria da Cunha
dc.creatorCâmara, Antonia Cláudia Jácome da
dc.creatorEspinoza, Bertha
dc.creatorNoya, Belkisyole Alarcón de
dc.creatorPuerta, Concepción
dc.creatorRiarte, Adelina
dc.creatorDiosque, Patricio
dc.creatorSosa-Estani, Sergio
dc.creatorGuhl, Felipe
dc.creatorRibeiro, Isabela
dc.creatorAznar, Christine
dc.creatorBrito, Constança
dc.creatorYadón, Zaída Estela
dc.creatorSchijman, Alejandro G.
dc.date2016-03-01T14:37:34Z
dc.date2016-03-01T14:37:34Z
dc.date2015
dc.date.accessioned2023-09-26T22:51:51Z
dc.date.available2023-09-26T22:51:51Z
dc.identifierRAMIREZ, Juan Carlos; et al. Analytical Validation of Quantitative Real-Time PCR Methods for Quantification of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients. J Mol Diagn., v.17, n.5, p.605-615, Sept. 2015.
dc.identifier1525-1578
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/12920
dc.identifier10.1016/j.jmoldx.2015.04.010
dc.identifier1943-7811
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8884103
dc.descriptionAn 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.
dc.formatapplication/pdf
dc.languageeng
dc.publisherElsevier
dc.rightsopen access
dc.subjectAmostras de Sangue
dc.subjectChagas disease
dc.subjectTrypanosoma cruzi
dc.subjectQuantitative Real-Time PCR Methods
dc.subjectBlood Samples
dc.subjectReação em Cadeia da Polimerase
dc.subjectDoença de Chagas
dc.subjectTrypanosoma cruzi
dc.titleAnalytical Validation of Quantitative Real-Time PCR Methods for Quantification of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients
dc.typeArticle


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