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J Med Microbiol 52 (2003), 127-135; DOI: 10.1099/jmm.0.04923-0
© 2003 Society for General Microbiology
ISSN 0022-2615

DIAGNOSTICS, TYPING AND IDENTIFICATION

Low-stringency single specific primer PCR for identification of Leptospira

Marluce A. Assunção Oliveira1, Otávia L. Caballero4, Annamaria R. Vago2, Rudy A. Harskeerl5, Álvaro J. Romanha6, Sérgio D. J. Pena3, Andrew J. G. Simpson4 and Matilde Cota Koury1

1–3Departamento de Microbiologia1, Departamento de Morfologia2 and Departamento de Bioquímica e Imunologia3, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, CP 486, CEP: 31270-901, Belo Horizonte, Minas Gerais, Brazil 4Laboratory of Cancer Genetics, Ludwig Institute for Cancer Research, São Paulo, Brazil 5Department of Biomedical Research, Royal Tropical Institute, Amsterdam, The Netherlands 6Centro de Pesquisas ‘René Rachou’ – FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil

Correspondence Matilde Cota Koury kourymat{at}mono.icb.ufmg.br

Received 22 March 2002 Accepted 23 October 2002

Thirty-five Leptospira serovars from the species Leptospira interrogans, Leptospira borgpetersenii, Leptospira santarosai, Leptospira kirschneri, Leptospira weilii, Leptospira biflexa and Leptospira meyeri were characterized by the low-stringency single specific primer PCR (LSSP-PCR) technique. LSSP-PCR analysis was performed to detect DNA polymorphisms in a 285 bp DNA fragment amplified from genomic DNA with G1 and G2 selected primers. Similar LSSP-PCR profiles were obtained for serovars from the same genomic species, while serovars from non-related species produced distinct multiband patterns. Based on the data from sequence analysis, all genomic fragments amplified with G1 and G2 primers from distinct serovars of Leptospira were 285 bp in length, with nucleotide variation observed most frequently among different genomic species. The simplicity and accuracy of the LSSP-PCR technique were found to be suitable for identification of Leptospira species.






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