INVESTIGADORES
ETCHEVERRIA Analia Ines
artículos
Título:
Recommendations for the detection of Leptospira in urine by PCR
Autor/es:
LUCCHESI, PAULA M.A.; ARROYO, GUILLERMO H.; ETCHEVERRÍA, ANALÍA I.; PARMA, ALBERTO E.; SEIJO, ALFREDO C.
Revista:
REVISTA DA SOCIEDADE BRASILEIRA DE MEDICINA TROPICAL
Editorial:
Sociedade Brasileira de Medicina Tropical
Referencias:
Lugar: Uberaba- Brasil; Año: 2004 vol. 37 p. 131 - 134
ISSN:
0037-8682
Resumen:
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 immediatelyafter 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.