title: Genotyping of Trypanosoma cruzi DTUs and Trypanosoma rangeli genetic groups in experimentally infected Rhodnius prolixus by PCR-RFLP.
abstract: The specific detection and genetic typing of trypanosomes that infect humans, mammalian reservoirs, and vectors is crucial for diagnosis and epidemiology. We utilized a PCR-RFLP assay that targeted subunit II of cytochrome oxidase and 24Sα-rDNA to simultaneously detect and discriminate six Trypanosoma cruzi discrete typing units (DTUs) and two genetic groups of Trypanosoma rangeli (KP1+/KP1-) in intestinal contents of experimentally infected Rhodnius prolixus. The PCR assays showed that in 23 of 29 (79.4%) mixed infections with the six T. cruzi DTUs and mixed infections with individual DTUs and/or groups KP1+ and KP1-, both parasites were successfully detected. In six mixed infections that involved TcIII, the TcI, TcII, TcV, and TcVI DTUs predominated to the detriment of TcIII, indicating the selection of genetic groups. Interactions between different genetic groups and vectors may lead to genetic selection over TcIII. The elimination of this DTU by the immune system of the vector appears unlikely because TcIII was present in other mixed infections (TcIII/TcIV and TcIII/KP1+). Both molecular markers used in this study were sensitive and specific, demonstrating their usefulness in a wide geographical area where distinct genotypes of these two species are sympatric. Although the cellular and molecular mechanisms that are involved in parasite-vector interactions are still poorly understood, our results indicate a dynamic selection toward specific T. cruzi DTUs in R. prolixus during mixed genotype infections.
authors.affiliation: Departamento de Ciências Básicas da Saúde, Universidade Estadual de Maringá (UEM), Avenida Colombo, 5790, Jardim Universitário, Maringá, Paraná 87020-900, Brazil; Docente do curso de Biomedicina, Faculdade Integrado, Campo Mourão, Rodovia BR 158, KM 207, Campo Mourão, Paraná 87300-970, Brazil. Electronic address: amandarnsa@gmail.com.
authors.forename: Amanda R N
authors.initials: AR
authors.lastname: Sá
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Santa Catarina (UFSC), Campus João David Ferreira Lima, Trindade, Florianópolis, Santa Catarina 88040-900, Brazil. Electronic address: gremalaquias@hotmail.com.
authors.forename: Greicy B M
authors.initials: GB
authors.lastname: Dias
authors.affiliation: Departamento de Ciências Básicas da Saúde, Universidade Estadual de Maringá (UEM), Avenida Colombo, 5790, Jardim Universitário, Maringá, Paraná 87020-900, Brazil. Electronic address: karen_yu.ki@hotmail.com.
authors.forename: Karen Y
authors.initials: KY
authors.lastname: Kimoto
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Santa Catarina (UFSC), Campus João David Ferreira Lima, Trindade, Florianópolis, Santa Catarina 88040-900, Brazil. Electronic address: mario.steindel@ufsc.br.
authors.forename: Mário
authors.initials: M
authors.lastname: Steindel
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Santa Catarina (UFSC), Campus João David Ferreira Lima, Trindade, Florianópolis, Santa Catarina 88040-900, Brazil. Electronic address: edmundo.grisard@ufsc.br.
authors.forename: Edmundo C
authors.initials: EC
authors.lastname: Grisard
authors.affiliation: Departamento de Ciências Básicas da Saúde, Universidade Estadual de Maringá (UEM), Avenida Colombo, 5790, Jardim Universitário, Maringá, Paraná 87020-900, Brazil. Electronic address: mjotoledo@uem.br.
authors.forename: Max Jean O
authors.initials: MJ
authors.lastname: Toledo
authors.affiliation: Departamento de Ciências Básicas da Saúde, Universidade Estadual de Maringá (UEM), Avenida Colombo, 5790, Jardim Universitário, Maringá, Paraná 87020-900, Brazil. Electronic address: mlgomes@uem.br.
authors.forename: Mônica L
authors.initials: ML
authors.lastname: Gomes
mesh_terms: D000818:Animals; D001938:Brazil / Q000453:epidemiology; D014355:Chagas Disease / Q000635:transmission*; D003105:Colombia / Q000453:epidemiology; D005838:Genotype; D006801:Humans; D007303:Insect Vectors / Q000469:parasitology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012239:Rhodnius / Q000469:parasitology*; D014349:Trypanosoma cruzi / Q000235:genetics* / Q000302:isolation & purification; D057509:Trypanosoma rangeli / Q000235:genetics* / Q000302:isolation & purification
keywords: 24Sα rDNA; Experimental infection; RFLP-COII; Triatomines; Trypanosoma cruzi; Trypanosoma rangeli