title: Hemi-nested PCR and RFLP methodologies for identifying blood meals of the Chagas disease vector, Triatoma infestans.
abstract: Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted by hematophagous reduviid bugs within the subfamily Triatominae. These vectors take blood meals from a wide range of hosts, and their feeding behaviors have been used to investigate the ecology and epidemiology of T. cruzi. In this study we describe two PCR-based methodologies that amplify a fragment of the 16S mitochondrial rDNA, aimed to improve the identification of blood meal sources for Triatoma infestans: a.--Sequence analyses of two heminested PCRs that allow the identification of mammalian and avian species, and b.--restriction fragment length polymorphism (RFLP) analysis from the mammalian PCR to identify and differentiate multi-host blood meals. Findings from both methodologies indicate that host DNA could be detected and the host species identified in samples from laboratory reared and field collected triatomines. The implications of this study are two-fold. First, these methods can be used in areas where the fauna diversity and feeding behavior of the triatomines are unknown. Secondly, the RFLP method led to the identification of multi-host DNA from T. infestans gut contents, enhancing the information provided by this assay. These tools are important contributions for ecological and epidemiological studies of vector-borne diseases.
authors.affiliation: Centers for Disease Control and Prevention, Division of Parasitic Diseases and Malaria, Atlanta, Georgia, United States of America.
authors.forename: Dawn M
authors.initials: DM
authors.lastname: Roellig
authors.forename: Luis A
authors.initials: LA
authors.lastname: Gomez-Puerta
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Mead
authors.forename: Jesus
authors.initials: J
authors.lastname: Pinto
authors.forename: Jenny
authors.initials: J
authors.lastname: Ancca-Juarez
authors.forename: Maritza
authors.initials: M
authors.lastname: Calderon
authors.forename: Caryn
authors.initials: C
authors.lastname: Bern
authors.forename: Robert H
authors.initials: RH
authors.lastname: Gilman
authors.forename: Vitaliano A
authors.initials: VA
authors.lastname: Cama
mesh_terms: D000818:Animals; D014355:Chagas Disease / Q000469:parasitology; D004199:Disease Vectors; D005247:Feeding Behavior; D006801:Humans; D007303:Insect Vectors / Q000378:metabolism; D051379:Mice; D009893:Opossums; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000097:blood / Q000235:genetics; D051381:Rats; D017422:Sequence Analysis, DNA; D014224:Triatoma / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000235:genetics
chemical_list: D012336:RNA, Ribosomal, 16S
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