TY - JOUR
T1 - Direct Leishmania species typing in Old World clinical samples
T2 - Evaluation of 3 sensitive methods based on the heat-shock protein 70 gene
AU - Montalvo, Ana M.
AU - Fraga, Jorge
AU - El Safi, Sayda
AU - Gramiccia, Marina
AU - Jaffe, Charles L.
AU - Dujardin, Jean Claude
AU - Van der Auwera, Gert
N1 - Publisher Copyright:
© 2014 Elsevier Inc.
PY - 2014
Y1 - 2014
N2 - In the diagnosis of leishmaniasis, identification of the causative Leishmania species is relevant for treatment, prognosis, and epidemiology. Three new hsp70-based PCR variants were developed and recently validated on clinical samples from Peru, without the need for culturing. We evaluated their performance on 133 clinical samples (bone marrow, blood, buffy coat, lymph node aspirates, lesion biopsies) from 42 cutaneous and 56 visceral leishmaniasis patients and 35 negative cases, all from Old World countries (Italy, Sudan, Israel, and Tunisia). The 3 new PCRs were significantly more sensitive than those previously described for hsp70, and their respective restriction fragment analyses were more efficient for species identification. In 79% of the parasitologically confirmed positive samples, the species could be identified directly from sample DNA. This evaluation demonstrated that these new tools are globally applicable in different geographical, clinical, and sampling contexts, and they could become the reference method for identification of Leishmania species in clinical specimens.
AB - In the diagnosis of leishmaniasis, identification of the causative Leishmania species is relevant for treatment, prognosis, and epidemiology. Three new hsp70-based PCR variants were developed and recently validated on clinical samples from Peru, without the need for culturing. We evaluated their performance on 133 clinical samples (bone marrow, blood, buffy coat, lymph node aspirates, lesion biopsies) from 42 cutaneous and 56 visceral leishmaniasis patients and 35 negative cases, all from Old World countries (Italy, Sudan, Israel, and Tunisia). The 3 new PCRs were significantly more sensitive than those previously described for hsp70, and their respective restriction fragment analyses were more efficient for species identification. In 79% of the parasitologically confirmed positive samples, the species could be identified directly from sample DNA. This evaluation demonstrated that these new tools are globally applicable in different geographical, clinical, and sampling contexts, and they could become the reference method for identification of Leishmania species in clinical specimens.
KW - Clinical sample
KW - Direct PCR
KW - Hsp70
KW - Leishmania
KW - Species identification
KW - Typing
UR - http://www.scopus.com/inward/record.url?scp=84926226404&partnerID=8YFLogxK
U2 - 10.1016/j.diagmicrobio.2014.05.012
DO - 10.1016/j.diagmicrobio.2014.05.012
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C2 - 25038029
AN - SCOPUS:84926226404
SN - 0732-8893
VL - 80
SP - 35
EP - 39
JO - Diagnostic Microbiology and Infectious Disease
JF - Diagnostic Microbiology and Infectious Disease
IS - 1
ER -