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Open AccessArticle

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

1
Department Research and Development, Bernhard-Nocht-Institute for Tropical Medicine (BNITM), 20359 Hamburg, Germany
2
Actually Medical Mission Institute, 97074 Wuerzburg, Germany
3
Chair of Medical Informatics, Friedrich-Alexander University of Erlangen-Nürnberg, 91054 Erlangen, Germany
4
Department Health Advocacy, Organization Wiwa Yugumaiun Bunkuanarrua Tayrona (OWYBT), Valledupar 200001, Colombia
5
Department of Bioinformatics, Biocenter, Functional Genomics and Systems Biology Group, Julius-Maximilians University, 97070 Wuerzburg, Germany
*
Author to whom correspondence should be addressed.
J. Clin. Med. 2020, 9(5), 1517; https://doi.org/10.3390/jcm9051517
Received: 19 March 2020 / Revised: 11 May 2020 / Accepted: 14 May 2020 / Published: 18 May 2020
(This article belongs to the Section Infectious Diseases)
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. View Full-Text
Keywords: Chagas disease; Chagas diagnosis; Chagas monitoring; Chagas real time PCR; Trypanosoma cruzi Chagas disease; Chagas diagnosis; Chagas monitoring; Chagas real time PCR; Trypanosoma cruzi
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Kann, S.; Kunz, M.; Hansen, J.; Sievertsen, J.; Crespo, J.J.; Loperena, A.; Arriens, S.; Dandekar, T. Chagas Disease: Detection of Trypanosoma cruzi by a New, High-Specific Real Time PCR. J. Clin. Med. 2020, 9, 1517.

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