title: Identification of Old World Leishmania spp. by specific polymerase chain reaction amplification of cysteine proteinase B genes and rapid dipstick detection.
abstract: We used the cysteine proteinase B (cpb) gene family of the trypanosomatid genus Leishmania as a target to develop rapid, specific, and easy-to-use polymerase chain reaction (PCR) tests to discriminate Leishmania infantum, Leishmania donovani, Leishmania tropica, Leishmania aethiopica, and Leishmania major. Identification of all 5 Old World species and validation of intraspecies variability are features lacking in other species-specific PCRs. Amplicon analysis was done on agarose gels and was further simplified by using an oligochromatography dipstick to detect L. infantum and L. donovani products. Because the analytical sensitivity is lower than that of certain other species- and genus-specific PCRs, our assays are especially valuable for use on cultured isolates or directly on cryostabilates. As such, they can be implemented by research and health centers having access to culturing, DNA isolation, and PCR.
authors.affiliation: Department of Parasitology, Institute of Tropical Medicine Antwerp, 2000 Antwerp, Belgium.
authors.forename: Thierry
authors.initials: T
authors.lastname: Laurent
authors.forename: Gert
authors.initials: G
authors.lastname: Van der Auwera
authors.forename: Mallorie
authors.initials: M
authors.lastname: Hide
authors.forename: Pascal
authors.initials: P
authors.lastname: Mertens
authors.forename: Wilber
authors.initials: W
authors.lastname: Quispe-Tintaya
authors.forename: Stijn
authors.initials: S
authors.lastname: Deborggraeve
authors.forename: Simonne
authors.initials: S
authors.lastname: De Doncker
authors.forename: Thierry
authors.initials: T
authors.lastname: Leclipteux
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Bañuls
authors.forename: Philippe
authors.initials: P
authors.lastname: Büscher
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Dujardin
mesh_terms: D000818:Animals; D002845:Chromatography; D003546:Cysteine Endopeptidases / Q000235:genetics*; D017931:DNA Primers; D017125:Genes, Protozoan / Q000235:genetics*; D006801:Humans; D007891:Leishmania / Q000145:classification* / Q000201:enzymology / Q000235:genetics*; D007893:Leishmania donovani / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D018314:Leishmania infantum / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D018320:Leishmania major / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D007895:Leishmania tropica / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D003546:Cysteine Endopeptidases