Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints versus real-time PCR.
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...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3290571?pdf=render |
| _version_ | 1828332624712564736 |
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| author | Adenizar D Chagas-Junior Caroline L R da Silva Luciane Marieta Soares Cleiton S Santos Carlos D C M Silva Daniel A Athanazio Mitermayer G dos Reis Flávia W Cruz McBride Alan J A McBride |
| author_facet | Adenizar D Chagas-Junior Caroline L R da Silva Luciane Marieta Soares Cleiton S Santos Carlos D C M Silva Daniel A Athanazio Mitermayer G dos Reis Flávia W Cruz McBride Alan J A McBride |
| author_sort | Adenizar D Chagas-Junior |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T21:08:56Z |
| format | Article |
| id | doaj.art-9f4461fc47f441f989ddc0814cfd7924 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T21:08:56Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-9f4461fc47f441f989ddc0814cfd79242022-12-22T02:29:54ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0172e3271210.1371/journal.pone.0032712Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints versus real-time PCR.Adenizar D Chagas-JuniorCaroline L R da SilvaLuciane Marieta SoaresCleiton S SantosCarlos D C M SilvaDaniel A AthanazioMitermayer G dos ReisFlávia W Cruz McBrideAlan J A McBrideA 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.http://europepmc.org/articles/PMC3290571?pdf=render |
| spellingShingle | Adenizar D Chagas-Junior Caroline L R da Silva Luciane Marieta Soares Cleiton S Santos Carlos D C M Silva Daniel A Athanazio Mitermayer G dos Reis Flávia W Cruz McBride Alan J A McBride Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints versus real-time PCR. PLoS ONE |
| title | Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints versus real-time PCR. |
| title_full | Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints versus real-time PCR. |
| title_fullStr | Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints versus real-time PCR. |
| title_full_unstemmed | Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints versus real-time PCR. |
| title_short | Detection and quantification of Leptospira interrogans in hamster and rat kidney samples: immunofluorescent imprints versus real-time PCR. |
| title_sort | detection and quantification of leptospira interrogans in hamster and rat kidney samples immunofluorescent imprints versus real time pcr |
| url | http://europepmc.org/articles/PMC3290571?pdf=render |
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