Trypanosoma cruzi: Evaluation of PCR as a Laboratory Tool to Follow up the Evolution of Parasite Load
| Autor: | Fabiana Nabarro FERRAZ, Denise Lessa ALEIXO, Ana Paula GRUENDLING, Mônica Lúcia GOMES, Max Jean de Ornelas TOLEDO, Silvana Marques DE ARAÚJO |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: | |
| Zdroj: | Iranian Journal of Parasitology, Vol 11, Iss 3 (2016) |
| Druh dokumentu: | article |
| ISSN: | 1735-7020 2008-238X |
| Popis: | Background: The study evaluated qualitative PCR, primers 121-122 as a tool to follow up evolution parasite load of Trypanosoma cruzi. Methods: The study was conducted at the State University of Maringa, in 2015. Step 1, dilutions 1/10 were performed from T. cruzi-Y strain to obtain preparations of 50,000-0.05 parasites/mL from which DNA were extracted, quantified, and amplified. Step 2, the extracted DNA in the dilutions 5-0.05 parasites/mL was re-diluted 1/10, 1/100, 1/1000, quantified, and amplified. Polyacrylamide gels were photographed and thicknesses of the 330 bp kDNA fragments were measured. Results: Step 1, in the dilutions 50,000-50 parasites/mL kDNA fragments had same thickness and, dilutions 5-0.05 parasites/mL showed progressive decrease in thicknesses and staining intensity of the 330 bp fragments. Step 2, demonstrated that dilutions of five (re-dilutions 1/10 and 1/100) and 0.5 (1/10) parasites/mL produced similar thicknesses of the 330 bp fragments obtained in Step 1. However, very dilute DNA samples make difficult to reproduce the fragments thicknesses. Conclusion: PCR, despite its limitations, was able to detect progressive decrease in thicknesses/staining intensity of kDNA fragments in the dilutions 5-0.05 parasites/mL. Hence, has the potential to be used to follow-up evolution of parasite load, not by quantifying the number of parasites, but by dynamic evolution of the fragments thicknesses during etiological treatment. |
| Databáze: | Directory of Open Access Journals |
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