| Author | Finamore-Araujo, Paula | |
| Author | Fonseca, Gabriel Lucio Silva da | |
| Author | Vieira, Cecília Stahl | |
| Author | Castro, Daniele Pereira de | |
| Author | Moreira, Otacilio Cruz | |
| Access date | 2022-09-27T13:12:32Z | |
| Available date | 2022-09-27T13:12:32Z | |
| Document date | 2022 | |
| Citation | FINAMORE-ARAUJO, Paula etal. RNA as a feasible marker of Trypanosoma cruzi viability during the parasite interaction with the triatomine vector Rhodnius prolixus (Hemiptera, Triatominae). PLoS Negl. Trop. Dis. v. 16, n 7. 16 p. July 2022. | en_US |
| ISSN | 1935-2727 | en_US |
| URI | https://www.arca.fiocruz.br/handle/icict/54878 | |
| Sponsorship | This study was financed in part by the Coordination for the Improvement of Higher Education Personnel (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES) - Finance Code 001 (P.F.A). This work also received financial support from Inova Fiocruz (VPPCB-007-FIO-18-2-72), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Fundação Carlos Chagas Filho de Amparo à Pesquisa do Rio de Janeiro (FAPERJ). O.C.M. is a researcher fellow of CNPq1D (311539/2020-3) and FAPERJ (JCNE, E-26/203.031/2018). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. | en_US |
| Language | eng | en_US |
| Publisher | Public Library of Science | en_US |
| Rights | open access | |
| Title | RNA as a feasible marker of Trypanosoma cruzi viability during the parasite interaction with the triatomine vector Rhodnius prolixus (Hemiptera, Triatominae) | en_US |
| Type | Article | |
| DOI | 10.1371/journal.pntd.0010535 | |
| Abstract | A recurring question concerning Trypanosoma cruzi DNA detection/quantification is related
to the fact that DNA amplification, by itself, does not differentiate between viable or dead
parasites. On the other hand, RNA can be considered a potential molecular marker of patho gens viability. Herein, we developed a quantitative real-time PCR with reverse Transcription
(RT-qPCR) to quantify viable T. cruzi in artificially infected Rhodnius prolixus whilst evaluat ing differences between DNA and mRNA quantification along the insect midgut during 5, 9,
15 and 29 days after feeding. The RT-qPCR presented an improved performance with line arities ranging from 107 to 102 parasites equivalents and 3 to 0.0032 intestine unit equiva lents, and efficiencies of 100.3% and 102.8% for both T. cruzi and triatomine targets,
respectively. Comparing both RT-qPCR and qPCR, we confirmed that RNA is faster
degraded, no longer being detected at day 1 after parasite lysis, while DNA detection was
stable, with no decrease in parasite load over the days, even after parasite lysis. We also
observed statistical differences between the quantification of the parasite load by DNA and
by RNA on day 15 after feeding of experimentally infected R. prolixus. When assessing dif ferent portions of the digestive tract, by RT-qPCR, we could detect a statistically significant
reduction in the parasite amount in the anterior midgut. Oppositely, there was a statistically
significant increase of the parasite load in the hindgut. In conclusion, for this study parasite’s
viability in R. prolixus digestive tract were assessed targeting T. cruzi mRNA. In addition, dif ferences between DNA and RNA detection observed herein, raise the possibility that RNA is
a potential molecular viability marker, which could contribute to understanding the dynamics
of the parasite infection in invertebrate hosts. | en_US |
| Affilliation | Instituto Oswaldo Cruz. Laboratório de Virologia Molecular. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Instituto Oswaldo Cruz. Laboratório de Virologia Molecular. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil / Universidade Federal Fluminense. Niterói, RJ, Brasil. | en_US |
| Affilliation | Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Instituto Oswaldo Cruz. Laboratório de Virologia Molecular. Rio de Janeiro, RJ, Brasil. | en_US |
| Subject | Trypanosoma cruzi | en_US |
| Subject | Rhodnius prolixus | en_US |
| Subject | Triatominae | en_US |
| Subject | Real-Time PCR | en_US |
