title: [Standardization of conditions for PCR detection of Leishmania spp. DNA in sand flies (Diptera, Psychodidae)].
abstract: The correct identification of etiological agents in vector insects is crucial for epidemiological studies. Identification of flagellates in such vectors, usually by dissection of the digestive tract and microscopic observation of the contents as well as attempts at parasite isolation from insects in culture media, have proven operationally inadequate and with poor diagnostic specificity, since female sand flies are also hosts for other flagellates like Trypanosoma and Endotrypanum. Due to the efficiency and specificity of DNA target sequence amplification by polymerase chain reaction (PCR), the latter could be used to investigate the presence of Leishmania in sand flies, although the insects need to be properly stored and the Leishmania DNA extracted using appropriate methodology. This paper describes methodologies to standardize sand fly storage and Leishmania DNA extraction in such specimens as a more practical method in field studies.
authors.affiliation: Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, Brasil. byapaiva@usp.br
authors.forename: Byanca Regina
authors.initials: BR
authors.lastname: de Paiva
authors.forename: Nagilá Francinete Costa
authors.initials: NF
authors.lastname: Secundino
authors.forename: Paulo Fillemon Paulocci
authors.initials: PF
authors.lastname: Pimenta
authors.forename: Eunice Aparecida Biacnhi
authors.initials: EA
authors.lastname: Galati
authors.forename: Heitor Franco
authors.initials: HF
authors.lastname: Andrade Junior
authors.forename: Rosely dos Santos
authors.initials: Rdos S
authors.lastname: Malafronte
mesh_terms: D000818:Animals; D006224:Cricetinae; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D007303:Insect Vectors / Q000469:parasitology*; D007891:Leishmania / Q000235:genetics* / Q000302:isolation & purification; D007896:Leishmaniasis / Q000635:transmission*; D051379:Mice; D015336:Molecular Probe Techniques / Q000592:standards; D016133:Polymerase Chain Reaction / Q000592:standards*; D011309:Preservation, Biological; D011576:Psychodidae / Q000469:parasitology*; D012680:Sensitivity and Specificity
chemical_list: D016054:DNA, Protozoan