DNA content analysis allows discrimination between Trypanosoma cruzi and Trypanosoma rangeli

• Published in: PloS one Vol. 12; no. 12; p. e0189907
• Main Authors: Naves, Lucila Langoni, da Silva, Marcos Vinícius, Fajardo, Emanuella Francisco, da Silva, Raíssa Bernardes, De Vito, Fernanda Bernadelli, Rodrigues, Virmondes, Lages-Silva, Eliane, Ramírez, Luis Eduardo, Pedrosa, André Luiz
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 19.12.2017; Public Library of Science (PLoS)
• Subjects: Biology and Life Sciences; Cell cycle; Cell lines; Chagas disease; Cloning; Content analysis; Cytometry; Deoxyribonucleic acid; DNA; Flow cytometry; Gene sequencing; Genetic aspects; Genomes; Leishmania major; Methods; Nucleotide sequence; Parasites; Parasitic diseases; Protozoa; Qualitative research; Research and Analysis Methods; Studies; Trypanosoma; Trypanosoma cruzi; Trypanosoma rangeli; Vector-borne diseases; Vectors
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0189907

Trypanosoma cruzi, a human protozoan parasite, is the causative agent of Chagas disease. Currently the species is divided into six taxonomic groups. The genome of the CL Brener clone has been estimated to be 106.4-110.7 Mb, and DNA content analyses revealed that it is a diploid hybrid clone. Trypanosoma rangeli is a hemoflagellate that has the same reservoirs and vectors as T. cruzi; however, it is non-pathogenic to vertebrate hosts. The haploid genome of T. rangeli was previously estimated to be 24 Mb. The parasitic strains of T. rangeli are divided into KP1(+) and KP1(-). Thus, the objective of this study was to investigate the DNA content in different strains of T. cruzi and T. rangeli by flow cytometry. All T. cruzi and T. rangeli strains yielded cell cycle profiles with clearly identifiable G1-0 (2n) and G2-M (4n) peaks. T. cruzi and T. rangeli genome sizes were estimated using the clone CL Brener and the Leishmania major CC1 as reference cell lines because their genome sequences have been previously determined. The DNA content of T. cruzi strains ranged from 87,41 to 108,16 Mb, and the DNA content of T. rangeli strains ranged from 63,25 Mb to 68,66 Mb. No differences in DNA content were observed between KP1(+) and KP1(-) T. rangeli strains. Cultures containing mixtures of the epimastigote forms of T. cruzi and T. rangeli strains resulted in cell cycle profiles with distinct G1 peaks for strains of each species. These results demonstrate that DNA content analysis by flow cytometry is a reliable technique for discrimination between T. cruzi and T. rangeli isolated from different hosts.