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Evaluation of Four Single-Locus Markers for Leishmania Species Discrimination by Sequencing

Gert Van der Auwera, Christophe Ravel, Jaco J. Verweij, Aldert Bart, Gabriele Schöniane and Ingrid Felger


Several genetic markers have been described for discriminating Leishmania species. In most reported cases, one or a few polymorphisms are the basis of species identification, and the methods were validated on a limited number of strains from a particular geographical region. Therefore, most techniques may underestimate the global intraspecies variability and are applicable only in certain areas. In addition, interlaboratory standardization is mostly absent, complicating comparisons among different studies. Here, we compared species typing results from all sequence polymorphisms found in four popular markers that can be applied directly on clinical samples: the miniexon or spliced leader, the internal transcribed spacer of the ribosomal DNA array, the 7SL RNA gene, and the heat shock protein 70 gene. Clustering was evaluated among 74 Leishmania strains, selected to represent a wide geographic distribution and genetic variability of the medically relevant species of the genus. Results were compared with a multilocus sequence typing (MLST) approach using 7 single-copy household genes and with multilocus enzyme electrophoresis (MLEE), still considered the gold standard by some. We show that strain groupings are highly congruent across the four different single-locus markers, MLST, and MLEE. Overall, the heat shock protein 70 gene and the miniexon presented the best resolutions for separating medically relevant species. As gene sequence analysis is validated here on a global scale, it is advocated as the method of choice for use in genetic, clinical, and epidemiological studies and for managing patients with unknown origins of infection, especially in Western infectious disease clinics dealing with imported leishmaniasis.

 J. Clin. Microbiol. 2014 52(4) 1098-1104


LeishMan recommendations for treatment of cutaneous and mucosal leishmaniasis in travelers, 2014.

Blum J, Buffet P, Visser L, Harms G, Bailey MS, Caumes E, Clerinx J, van Thiel PP, Morizot G, Hatz C, Dorlo TP and Lockwood DN.


Treatment of cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) in travelers is still controversial. Over the last decade, national and international consortia have published recommendations for treating CL in travelers. These guidelines harmonize many issues, but there are some discrepancies.
Leishmania parasites causing CL can now be genotyped by polymerase chain reaction techniques for detecting Leishmania DNA. Therefore, treatment recommendations can now be species based rather than based on geographical exposure. To review the evidence on which the recommendations were based, “LeishMan” (Leishmaniasis Management), a group of experts from 13 institutions in eight European countries, performed a PubMed MEDLINE) literature search and considered unpublished evidence and the experts’ own personal experiences. The Oxford evidence grading system was used to evaluate the information.
In this article, the authors provide practical treatment recommendations for imported CL and ML in Europe, drawn up from the review by the European experts.

 J Travel Med. 2014 21(2) 116-29.


Clinical aspects and management of cutaneous leishmaniasis in rheumatoid patients treated with TNF-α antagonists

Andreas L.C. Neumayr, Gloria Morizot, Leo G. Visser, Diana N.J. Lockwood, Bernhard R. Beck, Stefan Schneider, Guillaume Bellaud, Florence Cordoliani, Françoise Foulet, Emmanuel A. Laffitte, Pierre Buffet, Johannes A. Blum

Patients under immunosuppressive therapy with tumor necrosis factor alpha (TNF-α) antagonists are vulnerable to various opportunistic infections including leishmaniasis. We present a case series of 8 travellers developing cutaneous leishmaniasis whilst on TNF-α antagonist treatment and review the literature on aspects of cutaneous leishmaniasis developing in patients treated with TNF-α antagonists.

We make interim recommendations regarding the drug therapy used to maintain remission in travellers with rheumatoid disease travelling to leishmania prone areas. Despite having a medical condition requiring continued rheumatological review the interval to diagnosis appears not to be reduced compared to that described in non-rheumatoid patients. Rheumatologists and family doctors should be aware of the need for post-travel surveillance for leishmaniasis in rheumatoid patients on TNF-alpha antagonist treatment.

 Travel Medicine and Infectious Disease 2013 11(6) 412–420


Heat-shock protein 70 gene sequencing for Leishmania species typing in European tropical infectious disease clinics. 2013. Eurosurveillance 18 (30)

Leishmania species determination on clinical samples on the basis of partial sequencing of the heat-shock protein 70 gene (hsp70), without the need for parasite isolation

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Imported leishmaniasis in the Netherlands from 2005 to 2012: epidemiology, diagnostic techniques and sequence-based species typing from 195 patients. 2013. Eurosurveillance, 18(30).

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Local or systemic treatment for new world cutaneous leishmaniasis? re-evaluating the evidence for the risk of mucosal leishmaniasis. 2012. International Health. 4:153-163. 

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Presentations at Worldleish 5: World Congress on leishmaniosis 2013

A European consortium for the harmonisation of diagnosis, parasite typing and treatment of patients with leishmaniasis

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Systematic comparison of five genotypic markers for species discrimination in leishmania

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Developing evidence based clinical recommendations for the treatment of cutaneous leishmaniasis in Europe.

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