title: Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints versus real-time PCR.
abstract: A major limitation in the clinical management and experimental research of leptospirosis is the poor performance of the available methods for the direct detection of leptospires. In this study, we compared real-time PCR (qPCR), targeting the lipL32 gene, with the immunofluorescent imprint method (IM) for the detection and quantification of leptospires in kidney samples from the rat and hamster experimental models of leptospirosis. Using a virulent strain of Leptospira interrogans serovar Copenhageni, a chronic infection was established in the rat model, which were euthanized 28 days post-infection, while the hamster model simulated an acute infection and the hamsters were euthanized eight days after inoculation. Leptospires in the kidney samples were detected using culture isolation, qPCR and the IM, and quantified using qPCR and the IM. In both the acute and chronic infection models, the correlation between quantification by qPCR and the IM was found to be positive and statistically significant (P<0.05). Therefore, this study demonstrates that the IM is a viable alternative for not only the detection but also the quantification of leptospires, particularly when the use of qPCR is not feasible.
authors.affiliation: Gonçalo Moniz Institute, Oswaldo Cruz Foundation, Ministry of Health, Salvador, Bahia, Brazil.
authors.forename: Adenizar D
authors.initials: AD
authors.lastname: Chagas-Junior
authors.forename: Caroline L R
authors.initials: CL
authors.lastname: da Silva
authors.forename: Luciane Marieta
authors.initials: LM
authors.lastname: Soares
authors.forename: Cleiton S
authors.initials: CS
authors.lastname: Santos
authors.forename: Carlos D C M
authors.initials: CD
authors.lastname: Silva
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Athanazio
authors.forename: Mitermayer G
authors.initials: MG
authors.lastname: dos Reis
authors.forename: Flávia W Cruz
authors.initials: FW
authors.lastname: McBride
authors.forename: Alan J A
authors.initials: AJ
authors.lastname: McBride
mesh_terms: D000818:Animals; D006224:Cricetinae; D005260:Female; D007239:Infections; D007668:Kidney / Q000378:metabolism*; D007921:Leptospira interrogans / Q000378:metabolism*; D007922:Leptospirosis / Q000382:microbiology* / Q000473:pathology; D008647:Mesocricetus; D008856:Microscopy, Fluorescence / Q000379:methods*; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA