Biotinylated probes to detect Leptospira interrogans on dot blot hybridization or by in situ hybridization |
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Authors: | P. Fach Daniele Trap J.P. Guillou |
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Affiliation: | Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire Central de Recherches Vétérinaires, Maisons-Alfort, France. |
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Abstract: | Total genomic biotinylated probes which can identify leptospires by hybridization on filters or by in situ hybridization are described in this study. According to the weak G + C content of the strains studied (35-39%) and owing to the decreasing melting temperature (Tm) due to overbiotinylation, hybridization and wash temperatures were optimized at 33 degrees C and at 42 degrees C respectively. Fourteen serovars of Leptospira interrogans belonging to 11 different serogroups and three serovars of Leptospira biflexa were used in this study. Cross-hybridization results show that it is possible, by means of such probes, specifically to recognize pathogenic strains. These probes did not hybridize with the three saprophytic strains: L. buenos-aires, L. patoc and L. andamana. We also ran a total genomic probe, specific to the serovar buenos-aires which hybridizes only with homologous DNA. |
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