| Author | Muñoz-Calderón, Arturo | |
| Author | Gomess, Natalia Lins Silva | |
| Author | Apodaca, Sofia | |
| Author | Noya, Belkisyolé Alarcón de | |
| Author | Díaz-Bello, Zoraida | |
| Author | Souza, Leticia Rocha Quintino | |
| Author | Costa, Alexandre Dias Tavares | |
| Author | Britto, Constança | |
| Author | Moreira, Otacilio Cruz | |
| Author | Schijman, Alejandro G. | |
| Access date | 2021-04-08T19:29:16Z | |
| Available date | 2021-04-08T19:29:16Z | |
| Document date | 2021 | |
| Citation | MUÑOZ-CALDERON, Arturo et al. Toward the Establishment of a Single Standard Curve for Quantification of Trypanosoma cruzi Natural Populations Using a Synthetic Satellite Unit DNA Sequence. The Journal of Molecular Diagnostics, 13p, 2021. | pt_BR |
| ISSN | 1525-1578 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/46584 | |
| Language | eng | pt_BR |
| Publisher | Elsevier | pt_BR |
| Rights | open access | |
| Subject in Portuguese | Trypanosoma cruzi | pt_BR |
| Subject in Portuguese | Padrão único | pt_BR |
| Subject in Portuguese | Quantificação | pt_BR |
| Subject in Portuguese | Populações naturais | pt_BR |
| Subject in Portuguese | Unidade de Satélite Sintético | pt_BR |
| Subject in Portuguese | Uso | pt_BR |
| Title | Toward the Establishment of a Single Standard Curve for Quantification of Trypanosoma cruzi Natural Populations Using a Synthetic Satellite Unit DNA Sequence | pt_BR |
| Type | Article | |
| DOI | 10.1016/j.jmoldx.2021.01.007 | |
| Abstract | Accurate diagnostic tools and surrogate markers of parasitologic response to treatment are needed for
managing Chagas disease. Quantitative real-time PCR (qPCR) is used for treatment monitoring, but vari ability in copy dosage and sequences of molecular target genes among different Trypanosoma cruzi strains
limit the precision of quantitative measures. To improve qPCR quantification accuracy, we designed and
evaluated a synthetic DNA molecule containing a satellite DNA (satDNA) repeat unit as standard for
quantification of T. cruzi loads in clinical samples, independently of the parasite strain. Probit regression
analysis established for Dm28c (TcI) and CL-Brener (TcVI) stocks similar 95% limit of detection values [0.903
(0.745 to 1.497) and 0.667 (CI, 0.113 to 3.927) copy numbers/mL, respectively] when synthetic DNA was the
standard for quantification, allowing direct comparison of loads in samples infected with different discrete
typing units. This standard curve was evaluated in 205 samples (38 acute oral and 19 chronic Chagas disease
patients) from different geographical areas infected with various genotypes, including samples obtained
during treatment follow-up; high agreement with parasitic load trends using standard curves based on DNA
extracted from spiked blood with counted parasites was obtained. This qPCR-based quantification strategy
will be a valuable tool in phase 3 clinical trials, to follow up patients under treatment or at risk of reac tivation, and in experimental models using different parasite strains. | pt_BR |
| Affilliation | Laboratorio de Biología Molecular de la Enfermedad de Chagas. Instituto de Ingeniería Genética y Biología Molecular “Dr Héctor Torres” (INGEBI), Buenos Aires, Argentina. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Laboratorio de Biología Molecular de la Enfermedad de Chagas. Instituto de Ingeniería Genética y Biología Molecular “Dr Héctor Torres” (INGEBI), Buenos Aires, Argentina. | pt_BR |
| Affilliation | the Sección de Inmunologia. Instituto de Medicina Tropical “Dr Félix Pifano. Universidad Central de Venezuela, Caracas, Venezuela. | pt_BR |
| Affilliation | the Sección de Inmunologia,z Instituto de Medicina Tropical “Dr Félix Pifano,” Universidad Central de Venezuela, Caracas, Venezuela; | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidad Central de Venezuela, Caracas, Venezuela / Laboratório de Ciências e Tecnologias Aplicadas à Saúde (LaCTAS). Venezuela. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Laboratorio de Biología Molecular de la Enfermedad de Chagas. Instituto de Ingeniería Genética y Biología Molecular “Dr Héctor Torres” (INGEBI), Buenos Aires, Argentina. | pt_BR |
| Subject | Trypanosoma cruzi | pt_BR |
| Subject | Single Standard | pt_BR |
| Subject | Quantification | pt_BR |
| Subject | Natural Populations | pt_BR |
| Subject | Using | pt_BR |
| Subject | Synthetic Satellite Unit | pt_BR |
