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Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease

A loop-mediated isothermal amplification assay was evaluated as a surrogate marker of treatment failure in Chagas disease (CD). A convenience series of 18 acute or reactivated CD patients who received anti-parasitic treatment with benznidazole was selected—namely, nine orally infected patients: thre...

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Autores principales: Muñoz-Calderón, Arturo A., Besuschio, Susana A., Wong, Season, Fernández, Marisa, García Cáceres, Lady J., Giorgio, Patricia, Barcan, Laura A., Markham, Cole, Liu, Yanwen E., de Noya, Belkisyole Alarcón, Longhi, Silvia A., Schijman, Alejandro G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9142941/
https://www.ncbi.nlm.nih.gov/pubmed/35630354
http://dx.doi.org/10.3390/microorganisms10050909
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author Muñoz-Calderón, Arturo A.
Besuschio, Susana A.
Wong, Season
Fernández, Marisa
García Cáceres, Lady J.
Giorgio, Patricia
Barcan, Laura A.
Markham, Cole
Liu, Yanwen E.
de Noya, Belkisyole Alarcón
Longhi, Silvia A.
Schijman, Alejandro G.
author_facet Muñoz-Calderón, Arturo A.
Besuschio, Susana A.
Wong, Season
Fernández, Marisa
García Cáceres, Lady J.
Giorgio, Patricia
Barcan, Laura A.
Markham, Cole
Liu, Yanwen E.
de Noya, Belkisyole Alarcón
Longhi, Silvia A.
Schijman, Alejandro G.
author_sort Muñoz-Calderón, Arturo A.
collection PubMed
description A loop-mediated isothermal amplification assay was evaluated as a surrogate marker of treatment failure in Chagas disease (CD). A convenience series of 18 acute or reactivated CD patients who received anti-parasitic treatment with benznidazole was selected—namely, nine orally infected patients: three people living with HIV and CD reactivation, five chronic CD recipients with reactivation after organ transplantation and one seronegative recipient of a kidney and liver transplant from a CD donor. Fifty-four archival samples (venous blood treated with EDTA or guanidinium hydrochloride-EDTA buffer and cerebrospinal fluid) were extracted using a Spin-column manual kit and tested by T. cruzi Loopamp kit (Tc-LAMP, index test) and standardized real-time PCR (qPCR, comparator test). Of them, 23 samples were also extracted using a novel repurposed 3D printer designed for point-of-care DNA extraction (PrintrLab). The agreement between methods was estimated by Cohen’s kappa index and Bland–Altman plot analysis. The T. cruzi Loopamp kit was as sensitive as qPCR for detecting parasite DNA in samples with parasite loads higher than 0.5 parasite equivalents/mL and infected with different discrete typing units. The agreement between qPCR and Tc-LAMP (Spin-column) or Tc-LAMP (PrintrLab) was excellent, with a mean difference of 0.02 [CI = −0.58–0.62] and −0.04 [CI = −0.45–0.37] and a Cohen’s kappa coefficient of 0.78 [CI = 0.60–0.96] and 0.90 [CI = 0.71 to 1.00], respectively. These findings encourage prospective field studies to validate the use of LAMP as a surrogate marker of treatment failure in CD.
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spelling pubmed-91429412022-05-29 Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease Muñoz-Calderón, Arturo A. Besuschio, Susana A. Wong, Season Fernández, Marisa García Cáceres, Lady J. Giorgio, Patricia Barcan, Laura A. Markham, Cole Liu, Yanwen E. de Noya, Belkisyole Alarcón Longhi, Silvia A. Schijman, Alejandro G. Microorganisms Communication A loop-mediated isothermal amplification assay was evaluated as a surrogate marker of treatment failure in Chagas disease (CD). A convenience series of 18 acute or reactivated CD patients who received anti-parasitic treatment with benznidazole was selected—namely, nine orally infected patients: three people living with HIV and CD reactivation, five chronic CD recipients with reactivation after organ transplantation and one seronegative recipient of a kidney and liver transplant from a CD donor. Fifty-four archival samples (venous blood treated with EDTA or guanidinium hydrochloride-EDTA buffer and cerebrospinal fluid) were extracted using a Spin-column manual kit and tested by T. cruzi Loopamp kit (Tc-LAMP, index test) and standardized real-time PCR (qPCR, comparator test). Of them, 23 samples were also extracted using a novel repurposed 3D printer designed for point-of-care DNA extraction (PrintrLab). The agreement between methods was estimated by Cohen’s kappa index and Bland–Altman plot analysis. The T. cruzi Loopamp kit was as sensitive as qPCR for detecting parasite DNA in samples with parasite loads higher than 0.5 parasite equivalents/mL and infected with different discrete typing units. The agreement between qPCR and Tc-LAMP (Spin-column) or Tc-LAMP (PrintrLab) was excellent, with a mean difference of 0.02 [CI = −0.58–0.62] and −0.04 [CI = −0.45–0.37] and a Cohen’s kappa coefficient of 0.78 [CI = 0.60–0.96] and 0.90 [CI = 0.71 to 1.00], respectively. These findings encourage prospective field studies to validate the use of LAMP as a surrogate marker of treatment failure in CD. MDPI 2022-04-26 /pmc/articles/PMC9142941/ /pubmed/35630354 http://dx.doi.org/10.3390/microorganisms10050909 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Muñoz-Calderón, Arturo A.
Besuschio, Susana A.
Wong, Season
Fernández, Marisa
García Cáceres, Lady J.
Giorgio, Patricia
Barcan, Laura A.
Markham, Cole
Liu, Yanwen E.
de Noya, Belkisyole Alarcón
Longhi, Silvia A.
Schijman, Alejandro G.
Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease
title Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease
title_full Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease
title_fullStr Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease
title_full_unstemmed Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease
title_short Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease
title_sort loop-mediated isothermal amplification of trypanosoma cruzi dna for point-of-care follow-up of anti-parasitic treatment of chagas disease
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9142941/
https://www.ncbi.nlm.nih.gov/pubmed/35630354
http://dx.doi.org/10.3390/microorganisms10050909
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