Biotinylated probes to detect Leptospira interrogans on dot blot hybridization or by in situ hybridization.
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.
Fach
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P
Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire Central de Recherches Vétérinaires, Maisons-Alfort, France.
Trap
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D
Guillou
J P
JP
D001710:Biotin; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis*; D007921:Leptospira interrogans / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D011237:Predictive Value of Tests; D013045:Species Specificity
D015342:DNA Probes; D004269:DNA, Bacterial; D001710:Biotin