Recommendations for the detection of Leptospira in urine by PCR
Open Access
- 1 March 2004
- journal article
- Published by FapUNIFESP (SciELO) in Revista da Sociedade Brasileira de Medicina Tropical
- Vol. 37 (2) , 131-134
Abstract
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.Keywords
This publication has 13 references indexed in Scilit:
- Lethal Leptospiral Pulmonary Hemorrhage: An Emerging Disease in Buenos Aires, ArgentinaEmerging Infectious Diseases, 2002
- Formas graves de leptospirose: aspectos clínicos, demográficos e ambientaisRevista da Sociedade Brasileira de Medicina Tropical, 2001
- LeptospirosisClinical Microbiology Reviews, 2001
- Epidemic Leptospirosis Associated with Pulmonary Hemorrhage—Nicaragua, 1995The Journal of Infectious Diseases, 1998
- Differentiation of pathogenic and non-pathogenic leptospires by means of the polymerase chain reactionRevista do Instituto de Medicina Tropical de São Paulo, 1997
- Detection of seven species of pathogenic leptospires by PCR using two sets of primersJournal of General Microbiology, 1993
- Hemoptise/s e síndrome de angústia respiratória do adulto como causas de morte na leptospirose: mudanças de padrões clínicos e anatomopatológicosRevista da Sociedade Brasileira de Medicina Tropical, 1992
- Development of a PCR test specific for Leptospira hardjo genotype bovis1991
- Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA PolymeraseScience, 1988
- Differentiation of Pathogenic and Saprophytic Leptospires I. Growth at Low TemperaturesJournal of Bacteriology, 1967