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

• Published in: Memórias do Instituto Oswaldo Cruz Vol. 98; no. 3; pp. 373 - 376
• Main Authors: Lane, Joshua E, Ribeiro-Rodrigues, Rodrigo, Olivares-Villagómez, Danyvid, Vnencak-Jones, Cindy L, McCurley, Thomas L, Carter, Clint E
• Format: Journal Article
• Language: English; Portuguese
• Published: Brazil Instituto Oswaldo Cruz, Ministério da Saúde 01.04.2003; Fundação Oswaldo Cruz (FIOCRUZ)
• Subjects: Animals; direct in situ PCR; DNA, Kinetoplast - genetics; DNA, Kinetoplast - isolation & purification; DNA, Protozoan - genetics; DNA, Protozoan - isolation & purification; Heart - parasitology; In Situ Hybridization; in situ polymerase chain reaction (PCR); indirect in situ PCR; Mice; murine cardiac tissue; PARASITOLOGY; Polymerase Chain Reaction; TROPICAL MEDICINE; Trypanosoma cruzi; Trypanosoma cruzi - genetics; Trypanosoma cruzi - isolation & purification; indirect in situ PCR; direct in situ PCR; in situ polymerase chain reaction (PCR); murine cardiac tissue; Trypanosoma cruzi
• ISSN: 0074-0276; 1678-8060; 0074-0276; 1678-8060
• DOI: 10.1590/S0074-02762003000300013

The use of in situ techniques to detect DNA and RNA sequences has proven to be an invaluable technique with paraffin-embedded tissue. Advances in non-radioactive detection systems have further made these procedures shorter and safer. We report the detection of Trypanosoma cruzi, the causative agent of Chagas disease, via indirect and direct in situ polymerace chain reaction within paraffin-embedded murine cardiac tissue sections. The presence of three T. cruzi specific DNA sequences were evaluated: a 122 base pair (bp) sequence localized within the minicircle network, a 188 bp satellite nuclear repetitive sequence and a 177 bp sequence that codes for a flagellar protein. In situ hybridization alone was sensitive enough to detect all three T. cruzi specific DNA sequences.