Identification of Old World Leishmania spp. by specific polymerase chain reaction amplification of cysteine proteinase B genes and rapid dipstick detection.
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.
Laurent
Thierry
T
Department of Parasitology, Institute of Tropical Medicine Antwerp, 2000 Antwerp, Belgium.
Van der Auwera
Gert
G
Hide
Mallorie
M
Mertens
Pascal
P
Quispe-Tintaya
Wilber
W
Deborggraeve
Stijn
S
De Doncker
Simonne
S
Leclipteux
Thierry
T
Bañuls
Anne-Laure
AL
Büscher
Philippe
P
Dujardin
Jean-Claude
JC
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
D017931:DNA Primers; D003546:Cysteine Endopeptidases