Simone Kann1,2, Meik Kunz3, Jessica Hansen1, Jürgen Sievertsen1, Jose J Crespo4, Aristides Loperena4, Sandra Arriens1, Thomas Dandekar5. 1. Bernhard-Nocht-Institute for Tropical Medicine (BNITM), Department Research and Development, 20359 Hamburg, Germany. 2. Actually Medical Mission Institute, 97074 Wuerzburg, Germany. 3. Friedrich-Alexander University of Erlangen-Nürnberg, Chair of Medical Informatics, 91054 Erlangen, Germany. 4. Organization Wiwa Yugumaiun Bunkuanarrua Tayrona (OWYBT), Department Health Advocacy, 200001 Valledupar, Colombia. 5. Julius-Maximilians University, Department of Bioinformatics, Biocenter, Functional Genomics and Systems Biology Group, 97070 Wuerzburg, Germany.
Abstract
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.
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.
Entities:
Keywords:
Chagas diagnosis; Chagas disease; Chagas monitoring; Chagas real time PCR; Trypanosoma cruzi
Authors: Anou Dreyfus; Marie-Thérèse Ruf; Marga Goris; Sven Poppert; Anne Mayer-Scholl; Nadine Loosli; Nadja S Bier; Daniel H Paris; Tshokey Tshokey; John Stenos; Eliharintsoa Rajaonarimirana; Gustavo Concha; Jorge Orozco; Johana Colorado; Andrés Aristizábal; Juan C Dib; Simone Kann Journal: PLoS Negl Trop Dis Date: 2022-06-06