Fluorogenic assay for molecular typing of the Leishmania donovani complex: taxonomic and clinical applications.
We describe a new fluorogenic assay for the identification of species and intraspecies groups within the Leishmania donovani complex. The assay combined (1) 2 polymerase chain reactions targeting the 2 cysteine proteinase b isogenes and (2) a fluorescence-resonance energy transfer/melting curve analysis of the polymorphisms within a 31-nt region. All strains within the L. donovani complex were distinguished from L. tropica, L. major, and L. aethiopica, and 5 distinct groups were identified within the L. donovani complex. Discrepancies were observed with the present taxonomy on the basis of isoenzyme analysis and concerned East African strains, which suggests the need for a systematic reevaluation of the taxonomy. The capacity to type parasites directly from clinical samples was demonstrated with blood and bone marrow samples. This rapid and high-throughput alternative for molecular diagnosis and epidemiological studies of visceral leishmaniasis could be adapted for use with other Leishmania species.
Quispe-Tintaya
Kelly Wilber
KW
Laboratory of Molecular Parasitology, Prins Leopold Instituut voor Tropische Geneeskunde, Antwerp, Belgium.
Laurent
Thierry
T
Decuypere
Saskia
S
Hide
Mallorie
M
Bañuls
Anne-Laure
AL
De Doncker
Simonne
S
Rijal
Suman
S
Cañavate
Carmen
C
Campino
Lenea
L
Dujardin
Jean-Claude
JC
D000818:Animals; D003546:Cysteine Endopeptidases / Q000097:blood / Q000235:genetics*; D004283:Dog Diseases / Q000469:parasitology; D004285:Dogs; D005453:Fluorescence; D006801:Humans; D007891:Leishmania / Q000145:classification* / Q000201:enzymology / Q000235:genetics; D007898:Leishmaniasis, Visceral / Q000175:diagnosis / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000379:methods*; D015800:Protozoan Proteins / Q000097:blood / Q000235:genetics*; D013154:Spleen / Q000469:parasitology
D015800:Protozoan Proteins; D003546:Cysteine Endopeptidases