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Detection and identification of Leishmania spp.: application of two hsp70-based PCR-RFLP protocols to clinical samples from the New World

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Abstract

Leishmaniasis is highly prevalent in New World countries, where several methods are available for detection and identification of Leishmania spp. Two hsp70-based PCR protocols (PCR-N and PCR-F) and their corresponding restriction fragment length polymorphisms (RFLP) were applied for detection and identification of Leishmania spp. in clinical samples recruited in Colombia, Guatemala, and Honduras. A total of 93 cases were studied. The samples were classified into positive or suspected of leishmaniasis according to parasitological criteria. Molecular amplification of two different hsp70 gene fragments and further RFLP analysis for identification of Leishmania species was done. The detection in parasitologically positive samples was higher using PCR-N than PCR-F. In the total of samples studied, the main species identified were Leishmania panamensis, Leishmania braziliensis, and Leishmania infantum (chagasi). Although RFLP-N was more efficient for the identification, RFLP-F is necessary for discrimination between L. panamensis and Leishmania guyanesis, of great importance in Colombia. Unexpectedly, one sample from this country revealed an RFLP pattern corresponding to Leishmania naiffi. Both molecular variants are applicable for the study of clinical samples originated in Colombia, Honduras, and Guatemala. Choosing the better tool for each setting depends on the species circulating. More studies are needed to confirm the presence of L. naiffi in Colombian territory.

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Acknowledgements

The authors appreciate the contribution of the patients for kindly donating their biological samples to assess this investigation. The assistance of Honduran and Guatemalan health services for helping to PECET’S researchers in sampling collection is acknowledged. The valuable contribution of Horacio Cadenas (PECET) is also appreciated. This work was partially funded by the third framework program of the Belgian Directorate General for Development with ITM Antwerp. The scientific support of Jean-Claude Dujardin is deeply recognized.

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Authors and Affiliations

  1. Department of Parasitology, Institute of Tropical Medicine Pedro Kourí, Havana, Cuba

    Ana M. Montalvo, Jorge Fraga & Annia Alba

  2. PECET (Program of Study and Control of Tropical Diseases) University of Antioquia, Medellín, Colombia

    Dídier Tirado, Gustavo Blandón, Iván Darío Vélez & Carlos Muskus

  3. Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium

    Gert Van der Auwera

Authors
  1. Ana M. Montalvo
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  2. Jorge Fraga
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  4. Gustavo Blandón
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  5. Annia Alba
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  6. Gert Van der Auwera
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  7. Iván Darío Vélez
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  8. Carlos Muskus
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Contributions

AMM, JF, and CM conceived and designed the study. AMM, DT, GB performed the experiments. AMM, JF, AA, GVDA, and CM analyzed the data. AMM and GVDA wrote the paper.

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Correspondence to Ana M. Montalvo.

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Montalvo, A.M., Fraga, J., Tirado, D. et al. Detection and identification of Leishmania spp.: application of two hsp70-based PCR-RFLP protocols to clinical samples from the New World. Parasitol Res 116, 1843–1848 (2017). https://doi.org/10.1007/s00436-017-5454-6

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