Acta Tropica 78 (2001) 261–267 Use of DNA-based diagnostic methods for human leishmaniasis in Minas Gerais, Brazil Antero Silva Ribeiro de Andrade a, *, Rosa ˆngela Fa ´tima Gomes b , Octa ´vio Fernandes c , Maria Norma de Melo b a Comissa ˜o Nacional de Energia Nuclear /Centro de Desenolimento da Tecnologia Nuclear (CDTN/CNEN), Cidade Uniersita ´ria -Campus da UFMG, Rua Professor Ma ´rio Werneck, S /No, PO Box 941, Belo Horizonte, CEP 31120 -970 Minas Gerais, Brazil b Departamento de Parasitologia, Uniersidade Federal de Minas Gerais (UFMG), A. Anto ˆnio Carlos, 6627 Campus da UFMG, PO Box 486, Belo Horizonte, CEP 21270 -901 Minas Gerais, Brazil c Fundac ¸a ˜o Oswaldo Cruz, Departamento de Medicina Tropical (FIOCRUZ), A. Brazil 4365, CEP 21045 -900 Rio de Janeiro, Brazil Received 21 October 1999; received in revised form 16 November 2000; accepted 10 December 2000 Abstract DNA hybridisation was used to type 26 samples from lesions of human patients from the Rio Doce Valley (Minas Gerais, Brazil) clinically diagnosed as having cutaneous leishmaniasis, using kinetoplast DNA (kDNA) cloned mini-circle probes specific for the Leishmania mexicana and Leishmania braziliensis complexes. All samples were found to belong to the L. braziliensis complex. When biopsies were pressed directly onto touch blot membranes 38.5% of the samples were positive. The positivity and specificity obtained were both 100% when cultured blotted parasites were used. The results were confirmed by polymerase chain reaction (PCR) analysis using primers specific for the L. mexicana and L. braziliensis complexes. © 2001 Elsevier Science B.V. All rights reserved. Keywords: Leishmania ; Diagnosis; DNA hybridisation; PCR www.parasitology-online.com 1. Introduction New World leishmaniasis occurs from the southern United States to northern Argentina and is caused by 13 species grouped into three com- plexes, Leishmania (Viannia ) braziliensis, Leish - mania mexicana, and Leishmania donoani. Members of the L. braziliensis complex cause cutaneous or mucocutaneous lesions, those of the L. mexicana complex cause localised or diffuse cutaneous involvement and L. donoani complex species produce visceral disease (Grimaldi and Tesh, 1993). The tegumentary (cutaneous and mucocuta- neous) forms of leishmaniasis occur throughout Brazil. Prevalence of the disease has increased in * Corresponding author. Tel.: +55-31-4993182; fax: +55- 31-4993380. E-mail address: antero@urano.cdtn.br (A.S. Ribeiro de An- drade). 0001-706X/01/$ - see front matter © 2001 Elsevier Science B.V. All rights reserved. PII:S0001-706X(01)00081-X