| Literature DB >> 15094896 |
Paula M A Lucchesi1, Guillermo H Arroyo, Analía I Etcheverría, Alberto E Parma, Alfredo C Seijo.
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.Entities:
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Year: 2004 PMID： 15094896 DOI： 10.1590/s0037-86822004000200003
Source DB: PubMed Journal: Rev Soc Bras Med Trop ISSN： 0037-8682 Impact factor: 1.581