detection and quantification of leptospira interrogans in hamster and rat kidney samples immunofluorescent imprints versus real-time pcr检测和量化的钩端螺旋体interrogans仓鼠和老鼠肾脏样本immunofluorescent痕迹和实时pcr.pdfVIP

Detection and Quantification of Leptospira interrogans in Hamster and Rat Kidney Samples: Immunofluorescent Imprints versus Real-time PCR 1 1 2 1 Adenizar D. Chagas-Junior , Caroline L. R. da Silva , Luciane Marieta Soares , Cleiton S. Santos , 1 1,2 1 ´ 1 Carlos D. C. M. Silva , Daniel A. Athanazio , Mitermayer G. dos Reis , Flavia W. Cruz McBride , Alan J. A. McBride1,3* 1 Gonc¸alo Moniz Institute, Oswaldo Cruz Foundation, Ministry of Health, Salvador, Bahia, Brazil, 2 Health Sciences Institute, Federal University of Bahia, Salvador, Bahia, Brazil, 3 Biotechnology, Centre for Technology Development, Federal University of Pelotas, Pelotas, Rio Grande do Sul, Brazil 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