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Trypanosoma cruzi: Evaluation of PCR as a Laboratory Tool to Follow up the Evolution of Parasite Load
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 5...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Tehran University of Medical Sciences
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5256057/ https://www.ncbi.nlm.nih.gov/pubmed/28127346 |
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author | FERRAZ, Fabiana Nabarro ALEIXO, Denise Lessa GRUENDLING, Ana Paula GOMES, Mônica Lúcia TOLEDO, Max Jean de Ornelas DE ARAÚJO, Silvana Marques |
author_facet | FERRAZ, Fabiana Nabarro ALEIXO, Denise Lessa GRUENDLING, Ana Paula GOMES, Mônica Lúcia TOLEDO, Max Jean de Ornelas DE ARAÚJO, Silvana Marques |
author_sort | FERRAZ, Fabiana Nabarro |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5256057 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Tehran University of Medical Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-52560572017-01-26 Trypanosoma cruzi: Evaluation of PCR as a Laboratory Tool to Follow up the Evolution of Parasite Load FERRAZ, Fabiana Nabarro ALEIXO, Denise Lessa GRUENDLING, Ana Paula GOMES, Mônica Lúcia TOLEDO, Max Jean de Ornelas DE ARAÚJO, Silvana Marques Iran J Parasitol Short Communication 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. Tehran University of Medical Sciences 2016 /pmc/articles/PMC5256057/ /pubmed/28127346 Text en Copyright© Iranian Society of Parasitology & Tehran University of Medical Sciences This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly. |
spellingShingle | Short Communication FERRAZ, Fabiana Nabarro ALEIXO, Denise Lessa GRUENDLING, Ana Paula GOMES, Mônica Lúcia TOLEDO, Max Jean de Ornelas DE ARAÚJO, Silvana Marques Trypanosoma cruzi: Evaluation of PCR as a Laboratory Tool to Follow up the Evolution of Parasite Load |
title | Trypanosoma cruzi: Evaluation of PCR as a Laboratory Tool to Follow up the Evolution of Parasite Load |
title_full | Trypanosoma cruzi: Evaluation of PCR as a Laboratory Tool to Follow up the Evolution of Parasite Load |
title_fullStr | Trypanosoma cruzi: Evaluation of PCR as a Laboratory Tool to Follow up the Evolution of Parasite Load |
title_full_unstemmed | Trypanosoma cruzi: Evaluation of PCR as a Laboratory Tool to Follow up the Evolution of Parasite Load |
title_short | Trypanosoma cruzi: Evaluation of PCR as a Laboratory Tool to Follow up the Evolution of Parasite Load |
title_sort | trypanosoma cruzi: evaluation of pcr as a laboratory tool to follow up the evolution of parasite load |
topic | Short Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5256057/ https://www.ncbi.nlm.nih.gov/pubmed/28127346 |
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