Acta Tropica 74 (2000) 89–93 Characterization of Leishmania sp. strains isolated from autochthonous cases of human cutaneous leishmaniasis in Santa Catarina State, southern Brazil Edmundo C. Grisard a,e, *, Ma ´rio Steindel a , Jeffrey J. Shaw c , Edna A.Y. Ishikawa f , Carlos J. Carvalho-Pinto a , Iriane Eger-Mangrich a , Helena K. Toma d , Jose ´ H. Lima a , Alvaro J. Romanha b , David A. Campbell e a Departamento de Microbiologia e Parasitologia, CCB, Uniersidade Federal de Santa Catarina, Floriano ´polis, SC, Brazil b Laborato ´rio de Parasitologia Celular e Molecular -Centro de Pesquisas Rene ´ Rachou -FIOCRUZ, Belo Horizonte, MG, Brazil c Departamento de Parasitologia, ICB, Uniersidade de Sa ˜o Paulo, Sa ˜o Paulo, SP, Brazil d Departamento de Ana ´lises Clı ´nicas e Toxicolo ´gicas, CCS, Uniersidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil e Department of Microbiology and Immunology, UCLA School of Medicine, Los Angeles, CA, USA f Departamento de Parasitologia, Instituto Eandro Chagas -FNS, Bele ´m, PA, Brazil Received 30 June 1999; received in revised form 27 August 1999; accepted 9 September 1999 Abstract Four Leishmania sp. samples were isolated from autochthonous human cases of American cutaneous leishmaniasis (ACL) in Santa Catarina State, southern Brazil. These strains were characterized using indirect immunofluorescence with a panel of Leishmania -specific monoclonal antibodies (MAbs), and by PCR amplification and hybridization assay of the mini-exon gene with group specific probes. The results obtained with the MAbs were in agreement with the genetic marker. Two isolates (MHOM/BR/89/JSC89-H1 and MHOM/BR/89/JSC89-H2) were identified as L. (Leishmania ) amazonensis and two (MHOM/BR/96/LSC96-H3 and MHOM/BR/97/LSC97-H4) as L.(Viannia ) braziliensis. The southernmost autochthonous cases of ACL in Brazil are due to two different Leishmania sp. species, confirming the spreading of ACL on the American continent. © 2000 Elsevier Science B.V. All rights reserved. Keywords: Human cutaneous leishmaniasis; Leishmania sp.; Mini-exon gene; Spliced leader; Characterization; Monoclonal antibod- ies; PCR; Hybridization www.elsevier.com/locate/actatropica * Corresponding author. Tel.: +55-48-331-9512; fax: +55-48-331-9258. E-mail address: grisard@ccb.ufsc.br (E.C. Grisard) 0001-706X/00/$ - see front matter © 2000 Elsevier Science B.V. All rights reserved. PII:S0001-706X(99)00054-6