Fach P, Trap D, Guillou J P
Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire Central de Recherches Vétérinaires, Maisons-Alfort, France.
Lett Appl Microbiol. 1991 May;12(5):171-6. doi: 10.1111/j.1472-765x.1991.tb00532.x.
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.
本研究描述了可通过滤膜杂交或原位杂交来鉴定钩端螺旋体的全基因组生物素化探针。鉴于所研究菌株的G + C含量较低（35 - 39%），且由于过度生物素化导致解链温度（Tm）降低，杂交温度和洗涤温度分别优化为33℃和42℃。本研究使用了属于11个不同血清群的14个问号钩端螺旋体血清型和3个双曲钩端螺旋体血清型。交叉杂交结果表明，借助此类探针有可能特异性识别致病菌株。这些探针未与三种腐生菌株杂交：布宜诺斯艾利斯钩端螺旋体、帕托克钩端螺旋体和安达马纳钩端螺旋体。我们还制备了一种仅与同源DNA杂交的、针对布宜诺斯艾利斯血清型的全基因组探针。
