TY - JOUR
T1 - PCR diagnosis and characterization of Leishmania in local and imported clinical samples
AU - Schönian, Gabriele
AU - Nasereddin, Abedelmajeed
AU - Dinse, Nicole
AU - Schweynoch, Carola
AU - Schallig, Henk D. F. H.
AU - Presber, Wolfgang
AU - Jaffe, Charles L.
PY - 2003
Y1 - 2003
N2 - Leishmaniasis diagnosis in regions where multiple species exist should identify each species directly in the clinical sample without parasite culturing. The sensitivity of two PCR approaches which amplify part of the ssu rRNA gene and the ribosomal internal transcribed spacer (ITS), respectively, was determined using human and dog blood seeded with Leishmania promastigotes. ssu-rDNA-PCR was more sensitive than ITS1-PCR, however species identification was not possible by the former approach. When a nested ITS1-PCR was used its sensitivity equaled the ssu-rDNA-PCR. Digestion of ITS1 amplicon with the restriction enzyme HaeIII distinguished all medically relevant Leishmania species. ITS1-PCR was used to diagnose 162 local and imported suspected cases of leishmaniasis in Israel, the Palestinian Authority and Germany. 113 cases (69.7%) were positive by PCR and species identification was possible in 110 samples. Leishmania DNA was also amplified and identified at the species level from archived non-stained and Giemsa stained microscope slides
AB - Leishmaniasis diagnosis in regions where multiple species exist should identify each species directly in the clinical sample without parasite culturing. The sensitivity of two PCR approaches which amplify part of the ssu rRNA gene and the ribosomal internal transcribed spacer (ITS), respectively, was determined using human and dog blood seeded with Leishmania promastigotes. ssu-rDNA-PCR was more sensitive than ITS1-PCR, however species identification was not possible by the former approach. When a nested ITS1-PCR was used its sensitivity equaled the ssu-rDNA-PCR. Digestion of ITS1 amplicon with the restriction enzyme HaeIII distinguished all medically relevant Leishmania species. ITS1-PCR was used to diagnose 162 local and imported suspected cases of leishmaniasis in Israel, the Palestinian Authority and Germany. 113 cases (69.7%) were positive by PCR and species identification was possible in 110 samples. Leishmania DNA was also amplified and identified at the species level from archived non-stained and Giemsa stained microscope slides
U2 - https://doi.org/10.1016/S0732-8893(03)00093-2
DO - https://doi.org/10.1016/S0732-8893(03)00093-2
M3 - Article
C2 - 12967749
SN - 0732-8893
VL - 47
SP - 349
EP - 358
JO - Diagnostic microbiology and infectious disease
JF - Diagnostic microbiology and infectious disease
IS - 1
ER -