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Chagas Disease: Detection of Trypanosoma cruzi by a New, High-Specific Real Time PCR

Background: Chagas disease (CD) is a major burden in Latin America, expanding also to non-endemic countries. A gold standard to detect the CD causing pathogen Trypanosoma cruzi is currently not available. Existing real time polymerase chain reactions (RT-PCRs) lack sensitivity and/or specificity. We...

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Autores principales: Kann, Simone, Kunz, Meik, Hansen, Jessica, Sievertsen, Jürgen, Crespo, Jose J., Loperena, Aristides, Arriens, Sandra, Dandekar, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7291166/
https://www.ncbi.nlm.nih.gov/pubmed/32443464
http://dx.doi.org/10.3390/jcm9051517
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author Kann, Simone
Kunz, Meik
Hansen, Jessica
Sievertsen, Jürgen
Crespo, Jose J.
Loperena, Aristides
Arriens, Sandra
Dandekar, Thomas
author_facet Kann, Simone
Kunz, Meik
Hansen, Jessica
Sievertsen, Jürgen
Crespo, Jose J.
Loperena, Aristides
Arriens, Sandra
Dandekar, Thomas
author_sort Kann, Simone
collection PubMed
description Background: Chagas disease (CD) is a major burden in Latin America, expanding also to non-endemic countries. A gold standard to detect the CD causing pathogen Trypanosoma cruzi is currently not available. Existing real time polymerase chain reactions (RT-PCRs) lack sensitivity and/or specificity. We present a new, highly specific RT-PCR for the diagnosis and monitoring of CD. Material and Methods: We analyzed 352 serum samples from Indigenous people living in high endemic CD areas of Colombia using three leading RT-PCRs (k-DNA-, TCZ-, 18S rRNA-PCR), the newly developed one (NDO-PCR), a Rapid Test/enzyme-linked immuno sorbent assay (ELISA), and immunofluorescence. Eighty-seven PCR-products were verified by sequence analysis after plasmid vector preparation. Results: The NDO-PCR showed the highest sensitivity (92.3%), specificity (100%), and accuracy (94.3%) for T. cruzi detection in the 87 sequenced samples. Sensitivities and specificities of the kDNA-PCR were 89.2%/22.7%, 20.5%/100% for TCZ-PCR, and 1.5%/100% for the 18S rRNA-PCR. The kDNA-PCR revealed a 77.3% false positive rate, mostly due to cross-reactions with T. rangeli (NDO-PCR 0%). TCZ- and 18S rRNA-PCR showed a false negative rate of 79.5% and 98.5% (NDO-PCR 7.7%), respectively. Conclusions: The NDO-PCR demonstrated the highest specificity, sensitivity, and accuracy compared to leading PCRs. Together with serologic tests, it can be considered as a reliable tool for CD detection and can improve CD management significantly.
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spelling pubmed-72911662020-06-17 Chagas Disease: Detection of Trypanosoma cruzi by a New, High-Specific Real Time PCR Kann, Simone Kunz, Meik Hansen, Jessica Sievertsen, Jürgen Crespo, Jose J. Loperena, Aristides Arriens, Sandra Dandekar, Thomas J Clin Med Article Background: Chagas disease (CD) is a major burden in Latin America, expanding also to non-endemic countries. A gold standard to detect the CD causing pathogen Trypanosoma cruzi is currently not available. Existing real time polymerase chain reactions (RT-PCRs) lack sensitivity and/or specificity. We present a new, highly specific RT-PCR for the diagnosis and monitoring of CD. Material and Methods: We analyzed 352 serum samples from Indigenous people living in high endemic CD areas of Colombia using three leading RT-PCRs (k-DNA-, TCZ-, 18S rRNA-PCR), the newly developed one (NDO-PCR), a Rapid Test/enzyme-linked immuno sorbent assay (ELISA), and immunofluorescence. Eighty-seven PCR-products were verified by sequence analysis after plasmid vector preparation. Results: The NDO-PCR showed the highest sensitivity (92.3%), specificity (100%), and accuracy (94.3%) for T. cruzi detection in the 87 sequenced samples. Sensitivities and specificities of the kDNA-PCR were 89.2%/22.7%, 20.5%/100% for TCZ-PCR, and 1.5%/100% for the 18S rRNA-PCR. The kDNA-PCR revealed a 77.3% false positive rate, mostly due to cross-reactions with T. rangeli (NDO-PCR 0%). TCZ- and 18S rRNA-PCR showed a false negative rate of 79.5% and 98.5% (NDO-PCR 7.7%), respectively. Conclusions: The NDO-PCR demonstrated the highest specificity, sensitivity, and accuracy compared to leading PCRs. Together with serologic tests, it can be considered as a reliable tool for CD detection and can improve CD management significantly. MDPI 2020-05-18 /pmc/articles/PMC7291166/ /pubmed/32443464 http://dx.doi.org/10.3390/jcm9051517 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kann, Simone
Kunz, Meik
Hansen, Jessica
Sievertsen, Jürgen
Crespo, Jose J.
Loperena, Aristides
Arriens, Sandra
Dandekar, Thomas
Chagas Disease: Detection of Trypanosoma cruzi by a New, High-Specific Real Time PCR
title Chagas Disease: Detection of Trypanosoma cruzi by a New, High-Specific Real Time PCR
title_full Chagas Disease: Detection of Trypanosoma cruzi by a New, High-Specific Real Time PCR
title_fullStr Chagas Disease: Detection of Trypanosoma cruzi by a New, High-Specific Real Time PCR
title_full_unstemmed Chagas Disease: Detection of Trypanosoma cruzi by a New, High-Specific Real Time PCR
title_short Chagas Disease: Detection of Trypanosoma cruzi by a New, High-Specific Real Time PCR
title_sort chagas disease: detection of trypanosoma cruzi by a new, high-specific real time pcr
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7291166/
https://www.ncbi.nlm.nih.gov/pubmed/32443464
http://dx.doi.org/10.3390/jcm9051517
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