title: Detection of pathogenic leptospires in urine from naturally infected cattle by nested PCR.
abstract: A nested polymerase chain reaction (PCR) using primers from the LipL32 sequence of Leptospira spp. was used to detect shedding of pathogenic leptospires in urine from naturally infected cattle. Amplicons (497bp) were obtained from 21 pathogenic reference serovars belonging to four species (L. interrogans, L. borgpetersenii, L. santarosai, L. kirschneri). DNA was amplified from 26/30 urine samples taken from cattle with suspected leptospirosis and from leptospires cultivated from 10 of these samples. The limit of detection of DNA in the clinical samples was 200pg and the nested PCR detected all pathogenic reference serovars of Leptospira spp. tested. No PCR products were amplified using DNA from other common bacterial species from the bovine urogenital tract or urine, or from the non-pathogenic L. biflexa Andamana serovar. The nested PCR exhibited high specificity and sensitivity for detection of pathogenic serovars in urine from cattle.
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av Antônio Carlos, 6627, CP 486, CEP 31270-901, Belo Horizonte, Minas Gerais, Brazil. mrqbomfim@yahoo.com.br
authors.forename: Maria Rosa Quaresma
authors.initials: MR
authors.lastname: Bomfim
authors.forename: Edel Figueiredo
authors.initials: EF
authors.lastname: Barbosa-Stancioli
authors.forename: Matilde Cota
authors.initials: MC
authors.lastname: Koury
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology* / Q000652:urine*; D005260:Female; D007919:Leptospira / Q000302:isolation & purification*; D007922:Leptospirosis / Q000382:microbiology / Q000652:urine / Q000662:veterinary*; D014556:Urine / Q000382:microbiology