Recommendations for the detection of Leptospira in urine by PCR
In the present study PCR was applied to detect leptospires in human urine. Several approaches for sample processing were evaluated to optimize the detection of leptospires in urine mixed with this bacterium. Furthermore, some changes in the composition of the reaction mix were studied. No amplificat...
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| Format: | Article |
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Sociedade Brasileira de Medicina Tropical (SBMT)
2004-03-01
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| Series: | Revista da Sociedade Brasileira de Medicina Tropical |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822004000200003&tlng=en |
| _version_ | 1818548587106140160 |
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| author | Paula M.A. Lucchesi Guillermo H. Arroyo Analía I. Etcheverría Alberto E. Parma Alfredo C. Seijo |
| author_facet | Paula M.A. Lucchesi Guillermo H. Arroyo Analía I. Etcheverría Alberto E. Parma Alfredo C. Seijo |
| author_sort | Paula M.A. Lucchesi |
| collection | DOAJ |
| description | In the present study PCR was applied to detect leptospires in human urine. Several approaches for sample processing were evaluated to optimize the detection of leptospires in urine mixed with this bacterium. Furthermore, some changes in the composition of the reaction mix were studied. No amplification was observed in acidic urine, therefore neutralization of the sample immediately after collection is strongly recommended. PBS gave better results than Tris or NaOH as neutralizing reagents. Freezing and thawing of samples before processing yielded negative results. Elimination of epithelial cells, leukocytes and crystals by centrifugation at 3,000 rpm at room temperature increased sensitivity. In addition, both the washing step after collecting leptospires by centrifugation and the inclusion of 0.1% bovine serum albumin in the reaction mix minimized the interference of other inhibitory compounds. These modifications were useful to improve the detection of Leptospira in urine by PCR. |
| first_indexed | 2024-12-12T08:22:05Z |
| format | Article |
| id | doaj.art-804df5fc94fa4600a114530f2c5ffeda |
| institution | Directory Open Access Journal |
| issn | 1678-9849 |
| language | English |
| last_indexed | 2024-12-12T08:22:05Z |
| publishDate | 2004-03-01 |
| publisher | Sociedade Brasileira de Medicina Tropical (SBMT) |
| record_format | Article |
| series | Revista da Sociedade Brasileira de Medicina Tropical |
| spelling | doaj.art-804df5fc94fa4600a114530f2c5ffeda2022-12-22T00:31:22ZengSociedade Brasileira de Medicina Tropical (SBMT)Revista da Sociedade Brasileira de Medicina Tropical1678-98492004-03-01372131134Recommendations for the detection of Leptospira in urine by PCRPaula M.A. Lucchesi0Guillermo H. Arroyo1Analía I. Etcheverría2Alberto E. Parma3Alfredo C. Seijo4Universidad Nacional del Centro de la Provincia de Buenos AiresUniversidad Nacional del Centro de la Provincia de Buenos AiresUniversidad Nacional del Centro de la Provincia de Buenos AiresUniversidad Nacional del Centro de la Provincia de Buenos AiresHospital de Enfermedades Infecciosas 'Dr. Francisco Javier Muñiz'In the present study PCR was applied to detect leptospires in human urine. Several approaches for sample processing were evaluated to optimize the detection of leptospires in urine mixed with this bacterium. Furthermore, some changes in the composition of the reaction mix were studied. No amplification was observed in acidic urine, therefore neutralization of the sample immediately after collection is strongly recommended. PBS gave better results than Tris or NaOH as neutralizing reagents. Freezing and thawing of samples before processing yielded negative results. Elimination of epithelial cells, leukocytes and crystals by centrifugation at 3,000 rpm at room temperature increased sensitivity. In addition, both the washing step after collecting leptospires by centrifugation and the inclusion of 0.1% bovine serum albumin in the reaction mix minimized the interference of other inhibitory compounds. These modifications were useful to improve the detection of Leptospira in urine by PCR.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822004000200003&tlng=enLeptospiraPCRUrinePCR inhibitorsHuman |
| spellingShingle | Paula M.A. Lucchesi Guillermo H. Arroyo Analía I. Etcheverría Alberto E. Parma Alfredo C. Seijo Recommendations for the detection of Leptospira in urine by PCR Revista da Sociedade Brasileira de Medicina Tropical Leptospira PCR Urine PCR inhibitors Human |
| title | Recommendations for the detection of Leptospira in urine by PCR |
| title_full | Recommendations for the detection of Leptospira in urine by PCR |
| title_fullStr | Recommendations for the detection of Leptospira in urine by PCR |
| title_full_unstemmed | Recommendations for the detection of Leptospira in urine by PCR |
| title_short | Recommendations for the detection of Leptospira in urine by PCR |
| title_sort | recommendations for the detection of leptospira in urine by pcr |
| topic | Leptospira PCR Urine PCR inhibitors Human |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822004000200003&tlng=en |
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