title: Leishmania-FAST15: A rapid, sensitive and low-cost real-time PCR assay for the detection of Leishmania infantum and Leishmania braziliensis kinetoplast DNA in canine blood samples.
abstract: We describe an improved real-time PCR assay (designated as "Leishmania-FAST15") for the detection and quantification of Leishmania infantum and Leishmania braziliensis kinetoplast DNA minicircles in canine blood samples. The analytical sensitivity of this technique is 0.1 fg of DNA, which is equivalent to 0.002 parasite per reaction. This assay uses a small reaction volume (15 μl) and is rapid to perform, with the results being available in less than 34 min. This improved assay might also be suitable for detecting and quantifying L. infantum and L. braziliensis DNA in other tissues, such as bone marrow and lymph nodes.
authors.affiliation: Department of Immunology, Aggeu Magalhães Research Center, Recife, Brazil. Electronic address: filipe.dantas@cpqam.fiocruz.br.
authors.forename: Filipe
authors.initials: F
authors.lastname: Dantas-Torres
authors.affiliation: Department of Immunology, Aggeu Magalhães Research Center, Recife, Brazil.
authors.forename: Kamila Gaudêncio
authors.initials: KG
authors.lastname: da Silva Sales
authors.affiliation: Department of Genetics, Federal University of Pernambuco, Recife, Brazil.
authors.forename: Lidiane
authors.initials: L
authors.lastname: Gomes da Silva
authors.affiliation: Department of Veterinary Medicine, University of Bari, Valenzano, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Otranto
authors.affiliation: Department of Immunology, Aggeu Magalhães Research Center, Recife, Brazil.
authors.forename: Luciana Aguiar
authors.initials: LA
authors.lastname: Figueredo
mesh_terms: D000818:Animals; D003365:Costs and Cost Analysis; D018105:DNA, Kinetoplast / Q000097:blood* / Q000235:genetics; D004283:Dog Diseases / Q000097:blood / Q000235:genetics / Q000469:parasitology; D004285:Dogs / Q000097:blood* / Q000469:parasitology*; D007892:Leishmania braziliensis / Q000235:genetics / Q000302:isolation & purification*; D018314:Leishmania infantum / Q000235:genetics / Q000302:isolation & purification*; D018512:Parasitemia / Q000097:blood / Q000235:genetics / Q000469:parasitology; D060888:Real-Time Polymerase Chain Reaction / Q000191:economics*; D012015:Reference Standards; D012680:Sensitivity and Specificity
chemical_list: D018105:DNA, Kinetoplast
keywords: Blood; Diagnosis; Dogs; Leishmania spp.; Real-time PCR