Detection of Trypanosoma cruzi DNA within murine cardiac tissue sections by in situ polymerase chain reaction

Lane, Joshua E.; Ribeiro‐Rodrigues, Rodrigo; Olivares–Villagómez, Danyvid; McCurley, Thomas L.; Carter, Clint E.

doi:10.1590/s0074-02762003000300013

Cited by 12 publications

(8 citation statements)

References 20 publications

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“…Adad et al (1991) reported the finding of chronic inflammatory processes in the oesophagus of asymptomatic chronic chagasic patients and in those presenting megaoesophagus. These findings were confirmed by PCR, which amplified T. cruzi DNA both in the oesophagus of patients (Lages-Silva et al 2001, Elias et al 2003, Vago et al 2003 and in infected murine cardiac tissue (Lane et al 2003).…”

mentioning

confidence: 65%

Trypanosoma cruzi: parasite persistence in tissues in chronic chagasic Brazilian patients

Marcon

Albuquerque

Batista

et al. 2011

Mem. Inst. Oswaldo Cruz

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mentioning

confidence: 65%

Trypanosoma cruzi: parasite persistence in tissues in chronic chagasic Brazilian patients

Marcon

Albuquerque

Batista

et al. 2011

Mem. Inst. Oswaldo Cruz

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“…Regardless of the mode of infection or treatment regimen, the sacrifice of animals has typically been required to obtain information on dissemination of parasites and detection of parasites in specific tissues following infection. Furthermore, quantification of whole animal and organ-specific parasite burden has been both cumbersome and inconsistent, incorporating such techniques as PCR amplification or in situ hybridization of parasite-specific genes from tissue (Lane et al, 1997(Lane et al, , 2003Zhang and Tarleton, 1999;James et al, 2002), parasite antigen-specific immunofluoresence (Ben Younes-Chennoufi et al, 1988;Chandler and Watts, 1988;Taniwaki et al, 2007) and the counting of either nests of parasite amastigotes in tissue sections or free-swimming trypomastigotes in the blood (Nunes et al, 1990;Mortatti et al, 1992;Russo et al, 1996;Pinto et al, 1999). While these approaches have certainly been adequate for a variety of studies of experimental Chagas disease pathogenesis, they are also accompanied by significant limitations.…”

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confidence: 99%

Bioluminescent imaging of Trypanosoma cruzi infection

Hyland

Asfaw

Olson

et al. 2008

International Journal for Parasitology

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Chagas disease, caused by infection with the protozoan parasite Trypanosoma cruzi, is a major public health problem in Central and South America. The pathogenesis of Chagas disease is complex and the natural course of infection is not completely understood. The recent development of bioluminescence imaging technology has facilitated studies of a number of infectious and noninfectious diseases. We developed luminescent T. cruzi to facilitate similar studies of Chagas disease pathogenesis. Luminescent T. cruzi trypomastigotes and amastigotes were imaged in infections of rat myoblast cultures, which demonstrated a clear correlation of photon emission signal strength to the number of parasites used. This was also observed in mice infected with different numbers of luminescent parasites, where a stringent correlation of photon emission to parasite number was observed early at the site of inoculation, followed by dissemination of parasites to different sites over the course of a 25 day infection. Whole animal imaging from ventral, dorsal and lateral perspectives provided clear evidence of parasite dissemination. The tissue distribution of T. cruzi was further determined by imaging heart, spleen, skeletal muscle, lungs, kidneys, liver and intestines ex vivo. These results illustrate the natural dissemination of T. cruzi during infection and unveil a new tool for studying a number of aspects of Chagas disease, including rapid in vitro screening of potential therapeutic agents, roles of parasite and host factors in the outcome of infection, and analysis of differential tissue tropism in various parasite-host strain combinations.

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“…First, colonization of different tissues with T. cruzi described in patients and experimentally infected animals with organ damage 11,13 releases T. cruzi into the vascular system; these parasites then colonize other tissues. Second, infection is controlled by the immune system.…”

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confidence: 99%

Temporal Fluctuation of Infection with Different Trypanosoma cruzi Genotypes in the Wild Rodent Octodon degus

Campos¹,

Botto‐Mahan²,

Ortíz³

et al. 2010

The American Society of Tropical Medicine and Hygiene

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Chagas disease is a vector-borne zoonosis caused by the protozoa Trypanosoma cruzi . This taxon had been described as composed of two lineages (TCI and TCII) and five subgroups (IIa-IIe), but a recent study reported six lineages or discrete typing units (DTUs) ( T. cruzi I-VI).1 These lineages are defined as sets of stocks that are genetically more related to each other than to any other stock and are identifiable by common genetic molecular and immunologic markers. 2Trypanosoma cruzi populations circulate in nature in multiple T. cruzi genotypes that coexist in different hosts, including Octodon degus rodents.3-5 After a short acute or primary infection, the mammal host sustains subclinical infections, which are microscopically undetectable in peripheral blood during the undetermined and chronic phases. Conversely, parasitemia in those phases is detected only by polymerase chain reaction (PCR). The classic parasitologic diagnostic method for Chagas disease xenodiagnosis, which can amplify T. cruzi after feeding on infected hosts. 6 Although xenodiagnosis is specific, it lacks sensitivity and is limited to high levels of parasitemia.7 The epidemiology of Chagas disease and clinical symptoms are associated with the infective T. cruzi genotypes.8 Therefore, would be useful to know the dynamics of these genotypes.In the present study, we assess the occurrence of temporal fluctuations of T. cruzi DTUs in peripheral blood of two naturally infected wild reservoir specimens of O. degus by using a combination of two diagnosis methods: 1) xenodiagnosis with domestic and sylvatic vectors ( Triatoma infectans and Mepraia spinolai ), respectively, and 2) PCR DNA-based detection specific for minicircles and hybridization analyses with T. cruzi genotype-specific probes.Ten nymphs (stages II and III) of each vector species were allowed to feed simultaneously on anesthetized O. degus rodents for 30 minutes or until engorgement on the rodent (mean ± SD weight of ingested blood = 0.2 ± 0.05 mg). After 30 days, feces and intestinal contents of the triatomines were observed under a light microscope. The minimal theoretical parasitemia detected under these conditions is approximately 5 parasites/mL (1 parasite/0.2 mL). However, because several but not all insects (2-5) were parasite positive by visual examination, the estimated parasitemia would be > 10-25 parasites/mL.After microscopic inspection, the intestinal contents of each vector species pool was collected and PCR was performed as reported.9 Amplicons were subjected to electrophoresis on an agarose gel and transferred to nylon membranes. Copies of these membranes were hybridized separately with each probe under high stringency conditions.3 Construction of genotypespecific probes was performed as described.10 Different T. cruzi clones were used as templates to generate DNA probes to determine parasite genotypes. The probes were P 32 -labeled. 4A total of 35 O. degus were captured at the field and analyzed. Overall, only two O. degus showed infection with both vector species an...

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Detection of Trypanosoma cruzi DNA within murine cardiac tissue sections by in situ polymerase chain reaction

Cited by 12 publications

References 20 publications

Trypanosoma cruzi: parasite persistence in tissues in chronic chagasic Brazilian patients

Trypanosoma cruzi: parasite persistence in tissues in chronic chagasic Brazilian patients

Bioluminescent imaging of Trypanosoma cruzi infection

Temporal Fluctuation of Infection with Different Trypanosoma cruzi Genotypes in the Wild Rodent Octodon degus

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