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Diagn Microbiol Infect Dis. 2014 Sep;80(1):35-9. doi: 10.1016/j.diagmicrobio.2014.05.012. Epub 2014 May 21.

Direct Leishmania species typing in Old World clinical samples: evaluation of 3 sensitive methods based on the heat-shock protein 70 gene.

Author information

1
Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Havana, Cuba.
2
Faculty of Medicine, University of Khartoum, Sudan.
3
Unit of Vector-Borne Diseases and International Health, MIPI Department, Istituto Superiore di Sanità, Rome, Italy.
4
Kuvin Center for the Study of Infectious and Tropical Diseases, IMRIC, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
5
Department of BioMedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium; Department of BioMedical Sciences, University of Antwerp, Antwerp, Belgium.
6
Department of BioMedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium. Electronic address: gvdauwera@itg.be.

Abstract

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.

KEYWORDS:

Clinical sample; Direct PCR; Leishmania; Species identification; Typing; hsp70

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