Detection of Trypanosoma cruzi with the Polymerase Chain Reaction and in Situ Hybridization in Infected Murine Cardiac Tissue

Lane, Joshua E.; Olivares–Villagómez, Danyvid; Vnencak‐Jones, Cindy L.; McCurley, Thomas L.; Carter, C E

doi:10.4269/ajtmh.1997.56.588

Cited by 42 publications

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“…The demonstration of T. cruzi components in myocardium by more sensitive methods, such as immunohistochemistry and polymerase chain reaction (PCR), suggested that vestiges of parasites are necessary to trigger the inflammatory process (Higuchi et al 1993, Jones et al 1993, Bellotti et al 1996, Lane et al 1997, Olivares-Villagómez et al 1998. Jones et al (1993) also demonstrated that cardiac tissue adjacent to inflammatory infiltrates yielded PCR-amplified T. cruzi DNA in patients who died of severe chronic chagasic cardiomyopathy.…”

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confidence: 99%

Usefulness of PCR-based assays to assess drug efficacy in Chagas disease chemotherapy: value and limitations

Britto¹

2009

Mem. Inst. Oswaldo Cruz

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confidence: 99%

Usefulness of PCR-based assays to assess drug efficacy in Chagas disease chemotherapy: value and limitations

Britto¹

2009

Mem. Inst. Oswaldo Cruz

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“…Unlike liquid phase PCR, in situ detection allows precise localization of target sequences within tissue sections. In a previous report we demonstrated the efficacy of ISH to detect T. cruzi within cardiac tissue (Lane et al 1997).…”

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confidence: 93%

“…In the direct method, incorporation of the label during the elongation phase eliminates the need for subsequent hybridization. We previously described ISH to target three genes in order to identify T. cruzi within cardiac tissue (Lane et al 1997). This yielded a highly specific and sensitive method of detecting T. cruzi in tissue.…”

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confidence: 99%

“…The hearts were removed and embedded in paraffin. Primers and digoxigenin-labelled oligonucleotide probes used have been previously described (Lane et al 1997). Primers and probes specific for MCS, RS and FPS used in this study allowed detection of 0.167, 0.24, and 2.4 amastigote DNA equivalents, respectively (Lane et al 1997).…”

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confidence: 99%

“…In situ PCR was performed as described previously (Nuovo 1994, Lane et al 1997. To confirm the presence of T. cruzi amastigotes in the tissue, sections were stained with hematoxylin and eosin (H/E) and viewed under the microscope (Fig.…”

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

Lane

Ribeiro‐Rodrigues

Olivares–Villagómez

et al. 2003

Mem. Inst. Oswaldo Cruz

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Cornea as a tissue reservoir of Trypanosoma cruzi

et al. 2006

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Trypanosoma cruzi causal agent of Chagas' disease is a paninfective parasite of mammals transmitted through skin fecal contamination by Triatominae vectors. Studies of alternative routes for infection are scarce; therefore, eye infection should be important, because of the eye's high blood irrigation and brain proximity, as port of entry of the parasite. Trypanosoma cruzi parasites and/or their genetic material in ocular and adjacent muscle tissues were studied in batches of six NMRI mice (15 g) and Trichomys apereoides, an ancient caviomorph (250 g) inoculated with T. cruzi metacyclics from Brazilian (2) and Venezuelan (3) isolates genetically typified as T. cruzi I and II. Two animals/batch in the acute or chronic phase were killed and necropsies of cardiac and skeletal muscles, eyeball, and surrounding ocular muscle were processed for hematoxylin-eosine staining. Tissue parasitism was determined. DNA of the digested sections of the eyeball (5-10 mum) was extracted for T. cruzi k-DNA amplification by PCR, with S35 and S36 primers. The PCR products were analyzed. The average of maximum values of parasitemia of all infected animals was of 10(5) trypomastigotes/ml blood. Skeletal muscle and heart were colonized in patent infection for all isolates. Amastigote nests were found in corneal tissue of 2/3 of the used isolates and adjacent ocular muscle and connective tissue were parasitized. Trypanosoma cruzi k-DNA (330-bp band) was observed in ocular tissue of 4/6 of the isolates studied in both animal models. Investigations concerning infection of the eye globe tissues by T. cruzi are extremely scarce. The presence of stages of T. cruzi and/or its genetic products in ocular tissues indicate a broad colonization from a systemic infection. The results show the ocular environment as a possible appropriate microniche for T. cruzi and emphasize the risk of transmitting T. cruzi by ocular fluids and by parasitized cornea through transplants.

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Detection of Trypanosoma cruzi with the Polymerase Chain Reaction and in Situ Hybridization in Infected Murine Cardiac Tissue

Cited by 42 publications

References 0 publications

Usefulness of PCR-based assays to assess drug efficacy in Chagas disease chemotherapy: value and limitations

Usefulness of PCR-based assays to assess drug efficacy in Chagas disease chemotherapy: value and limitations

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

Cornea as a tissue reservoir of Trypanosoma cruzi

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