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Universal PCR assays for the differential detection of all Old World Leishmania species

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Abstract

For the epidemiological monitoring and clinical case management of leishmaniasis, determination of the causative Leishmania species gains importance. Current assays for the Old World often suffer from drawbacks in terms of validation on a geographically representative sample set and the ability to recognize all species complexes. We want to contribute to standardized species typing for Old World leishmaniasis. We determined the ribosomal DNA internal transcribed spacer 1 sequence of 24 strains or isolates, and validated four species-specific polymerase chain reactions (PCRs) amplifying this target. They discriminate L. aethiopica, L. tropica, L. major, and the L. donovani complex, use the same cycling conditions, and include an internal amplification control. Our PCRs amplify 0.1 pg of Leishmania DNA, while being 100% specific for species identification on an extensive panel of geographically representative strains and isolates. Similar results were obtained in an endemic reference laboratory in Kenya. Species could also be identified in clinical specimens. The presented PCRs require only agarose gel detection, and have several other advantages over many existing assays. We outline potential problems, suggest concrete solutions for transferring the technique to other settings, and deliver the proof-of-principle for analyzing clinical samples.

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Acknowledgments

We thank Gabi Schönian (Institut für Mikrobiologie und Hygiene, Berlin, Germany), Henk Schallig and Gerard Schoone (Koninklijk Instituut voor de Tropen, Amsterdam, The Netherlands), and Isabel Mauricio (London School of Hygiene and Tropical Medicine, London, United Kingdom) for contributing DNA from reference strains used in the study. Clinical samples were obtained from: (1) Lenea Campino (Institute of Hygiene and Tropical Medicine, Lisbon, Portugal); (2) Luigi Gradoni and Marina Grammicia (Istituto Superiore di Sanità, Rome, Italy); (3) Hechmi Louzir and Afif Ben Salah (Institut Pasteur de Tunis, Tunis, Tunisia); (4) Chantal Van Overmeir, Umberto d’Alessandro, Pim De Rijk, Leen Rigouts, Françoise Portaels, Stijn Deborggraeve, and Philippe Büscher (Institute of Tropical Medicine Antwerp, Antwerp, Belgium). This study was financed by the commission of the European Community’s Sixth Framework Programme, priority INCO-DEV, project TRYLEIDIAG, contract 015379. SOCO and GVdA are supported by the third framework program of the Belgian Development Cooperation with ITMA.

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Parasitology, Institute of Tropical Medicine Antwerp, Nationalestraat 155, 2000, Antwerp, Belgium

    S. Ogado Ceasar Odiwuor, S. De Doncker, I. Maes, P. Büscher, J.-C. Dujardin & G. Van der Auwera

  2. Center for Clinical Research, Kenya Medical Research Institute, Nairobi, Kenya

    S. Ogado Ceasar Odiwuor & M. Mbuchi

  3. Faculty of Medicine, University of Khartoum, Khartoum, Sudan

    A. Ageed Saad & S. El Safi

  4. Coris BioConcept, Gembloux, Belgium

    T. Laurent

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  1. S. Ogado Ceasar Odiwuor
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  2. A. Ageed Saad
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  3. S. De Doncker
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  4. I. Maes
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  5. T. Laurent
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  6. S. El Safi
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  7. M. Mbuchi
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  8. P. Büscher
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  9. J.-C. Dujardin
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  10. G. Van der Auwera
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Corresponding author

Correspondence to G. Van der Auwera.

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Ogado Ceasar Odiwuor, S., Ageed Saad, A., De Doncker, S. et al. Universal PCR assays for the differential detection of all Old World Leishmania species. Eur J Clin Microbiol Infect Dis 30, 209–218 (2011). https://doi.org/10.1007/s10096-010-1071-3

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  • DOI: https://doi.org/10.1007/s10096-010-1071-3

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