American tegumentary leishmaniasis: antigen-gene polymorphism, taxonomy and clinical pleomorphism A.L. Garcia a,b , A. Kindt b , K.W. Quispe-Tintaya b,c , H. Bermudez a , A. Llanos c , J. Arevalo c , A.L. Ban ˜uls d , S. De Doncker b , D. Le Ray b , J.C. Dujardin b, * a Centro Universitario de Medicina Tropical, Casilla 3119, Cochabamba, Bolivia b Laboratory of Molecular Parasitology, Instituut voor Tropische Geneeskunde ‘Prins Leopold’, 155 Nationalestraat, B-2000 Antwerpen, Belgium c Departamento de Bioquimica, Biologia Molecular y Farmacologia, Instituto de Medicina Tropical ‘Alexander von Humboldt’, Universidad Peruana Cayetano Heredia, A.P. 5045, Lima 100, Peru d Ge ´ne ´tique et Evolution des Maladies Infectieuses, UMR CNRS/IRD 2724, Montpellier, France Received 3 May 2004; received in revised form 1 July 2004; accepted 1 July 2004 Available online 24 August 2004 Abstract Multi-locus enzyme electrophoresis is the current gold standard for the genetic characterisation of Leishmania. However, this method is time-consuming and, more importantly, cannot be directly applied to parasites present in host tissue. PCR-based methods represent an ideal alternative but, to date, a multi-locus analysis has not been applied to the same sample. This has now been achieved with a sample of 55 neotropical isolates (Leishmania (Viannia) braziliensis, L. (V.) peruviana, L. (V.) guyanensis, L. (V.) lainsoni and L. (L.) amazonensis), using five different genes as targets, four of which encoded major Leishmania antigens (gp63, Hsp70, H2B and Cpb). Our multi-locus approach strongly supports the current taxonomy and demonstrates a highly robust method of distinguishing different strains. Within L. (V.) braziliensis, we did not encounter so far specific genetic differences between parasites isolated from cutaneous and mucosal lesions. Interestingly, results provided by each of the different antigen-genes in the species considered, were different, suggesting different selective pressures. Our work emphasises the need for a multi-disciplinary approach to study the clinical pleomorphism of leishmaniasis. # 2004 Elsevier B.V. All rights reserved. Keywords: Muco-cutaneous leishmaniasis; Multi-locus PCR; Antigens; Taxonomy 1. Introduction Leishmaniasis is endemic in 88 countries on four con- tinents, causing an estimated disease burden of 2,357,000 disability adjusted life years and 59,000 deaths per annum (WHO, 2002). This disease, caused by the protozoan para- site Leishmania, is characterised by a considerable clinical pleomorphism. Severity varies from self-limiting cutaneous lesions of the skin to severe mutilating mucosal manifesta- tions or fatal visceral forms. The paradigm of the disease is that specific clinical manifestations are generally associated with particular Leishmania species, but within a given species, different clinical forms can be encountered. This is typically illustrated by muco-cutaneous leishmaniasis— this severe syndrome encountered in Latin America has been reported in patients infected with Leishmania (Viannia) guyanensis and L. (V.) panamensis (Thomaz-Soccol et al., 2000), but is essentially caused by one species, L. (V.) braziliensis. Of all patients infected with L. (V.) braziliensis and having developed a primary cutaneous lesion, 10% will develop a metastatic mucosal presentation during their life- time (Campos, 1990). This is likely to be the results of interaction between host and parasite factors. With this in mind, parasite genotyping plays an essential role in both clinical and epidemiological risk assessment. Ideally, genetic characterisation tools should combine five features: (i) direct applicability on host tissues, (ii) easy, rapid and www.elsevier.com/locate/meegid Infection, Genetics and Evolution 5 (2005) 109–116 * Corresponding author. Tel.: +32-3-2476355; fax: +32-3-2476359. E-mail address: jcdujard@itg.be (J.C. Dujardin). 1567-1348/$ – see front matter # 2004 Elsevier B.V. All rights reserved. doi:10.1016/j.meegid.2004.07.003