[Standardization of conditions for PCR detection of Leishmania spp. DNA in sand flies (Diptera, Psychodidae)].
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.
de Paiva
Byanca Regina
BR
Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, Brasil. byapaiva@usp.br
Secundino
Nagilá Francinete Costa
NF
Pimenta
Paulo Fillemon Paulocci
PF
Galati
Eunice Aparecida Biacnhi
EA
Andrade Junior
Heitor Franco
HF
Malafronte
Rosely dos Santos
Rdos S
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
D016054:DNA, Protozoan