title: Combination of cytochrome b heteroduplex-assay and sequencing for identification of triatomine blood meals.
abstract: The identification of blood meals in vectors contributes greatly to the understanding of interactions between vectors, microorganisms and hosts. The aim of the current work was to complement the validation of cytochrome b (Cytb) heteroduplex assay (HDA) previously described, and to add the sequencing of the Cytb gene of some samples for the identification of blood meals in triatomines. Experimental feedings of reared triatomines helped to clarify the sensitivity of the HDA. Moreover, the sequencing coupled with the HDA, allowed the assessment of the technique's taxonomic level of discrimination. The primers used to produce DNA fragments of Cytb genes for HDA had a very high sensitivity for vertebrate DNAs, rather similar for mammals, birds and reptiles. However, the formation of heteroduplex depended on blood meal's quality rather than its quantity; a correlation was observed between blood meals' color and the positivity of HDA. HDA electrophoresis profiles were reproducible, and allowed the discrimination of blood origins at the species level. However, in some cases, intraspecific variability of Cytb gene generated different HDA profiles. The HDA based on comparison of electrophoresis profiles is a very useful tool for screening large samples to determine blood origins; the subsequent sequencing of PCR products of Cytb corresponding to different HDA profiles allowed the identification of species whatever the biotope in which the vectors were captured.
authors.affiliation: MIVEGEC (Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle), Université de Montpellier 1 et 2, CNRS 5290, IRD 224, Institut de Recherche pour le Développement (IRD), Representation in Bolivia, Av. H. Siles # 5290, CP 9214, La Paz, Bolivia. rosiob2002@yahoo.com
authors.forename: Rosio
authors.initials: R
authors.lastname: Buitrago
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Depickère
authors.forename: Marie-France
authors.initials: MF
authors.lastname: Bosseno
authors.forename: Edda Siñani
authors.initials: ES
authors.lastname: Patzi
authors.forename: Etienne
authors.initials: E
authors.lastname: Waleckx
authors.forename: Renata
authors.initials: R
authors.lastname: Salas
authors.forename: Claudia
authors.initials: C
authors.lastname: Aliaga
authors.forename: Simone Frédérique
authors.initials: SF
authors.lastname: Brenière
mesh_terms: D000818:Animals; D001769:Blood; D045303:Cytochromes b / Q000235:genetics*; D053938:DNA Fragmentation; D017931:DNA Primers; D020180:Heteroduplex Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D014225:Triatominae / Q000737:chemistry*
chemical_list: D017931:DNA Primers; D045303:Cytochromes b