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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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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. |
format | Online Article Text |
id | pubmed-7291166 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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