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Title: Molecular detection and parasite load of Trypanosoma cruzi in digestive tract tissue of Chagas disease patients affected by megacolon
Author: Pinto, Lilian
Schijman, Alejandro G.
Alonso Padilla, Julio
Lozano Beltran, Daniel Franz
Torrico, Mary Cruz
Gamba, Pietro
Torrez, Margarita
Lozada, Vania
Cartagena, Karina
Sánchez, Jareth
Torrico, Faustino
Gascón i Brustenga, Joaquim
Keywords: Malaltia de Chagas
Chagas' disease
Parasitologia mèdica
Medical parasitology
Issue Date: 1-Nov-2022
Abstract: Chagas disease, caused by the Trypanosoma cruzi parasite in the Americas affects similar to 7 million people, 30% with cardiac tissue damage and 10-15% with digestive disorders. In this study, we have developed a protocol to detect the presence of the parasite and estimate its load in resected dysfunctional tissue segments of chronically infected patients with digestive megacolon. We have included samples from 43 individuals, 38/5 with positive/negative serology for Chagas disease and digestive syndromes. Samples of 1.5 to 2.0 cm(2) were taken from different points of the dysfunctional digestive tract in specialized centres in Cochabamba, Bolivia. T. cruzi cultures were performed by inoculation with NNN-LIT culture medium, and genomic material was obtained from the samples for multiplex qPCR with TaqMan probes targeting satellite nuclear DNA. Cultures failed to isolate T. cruzi but qPCR reached a sensitivity of 42.1% (16/38) with all three spots and in triplicate. A new quantification methodology using synthetic satellite DNA as quantitation standard revealed parasite loads ranging from 2.2 x 10(2) to 1.0 x 10(6) satellite DNA copies/mu l. Positive samples from the distal end showed a higher parasite load. The results of the present study strengthen and add further evidence to previous findings in an experimental mouse model of chronic T. cruzi infection, providing a valuable tool to improve scientific knowledge on the relevance of the digestive tract in parasite persistence, and underline the need of a better understanding of host-pathogen interaction in digestive tissues, considering pathophysiology, disease immunology and response to treatment.
Note: Versió postprint del document publicat a:
It is part of: Acta Tropica, 2022, vol. 235
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ISSN: 1873-6254
Appears in Collections:Articles publicats en revistes (ISGlobal)

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