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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...

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Autores principales: FERRAZ, Fabiana Nabarro, ALEIXO, Denise Lessa, GRUENDLING, Ana Paula, GOMES, Mônica Lúcia, TOLEDO, Max Jean de Ornelas, DE ARAÚJO, Silvana Marques
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Tehran University of Medical Sciences 2016
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.
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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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