【摘要】 用泰泽氏菌RJ株感染的大鼠肝组织匀浆腹腔注射Scid小鼠和BALB/c小鼠,提取注射后小鼠肝、肾、肺、脾的DNA,PCR扩增,核酸杂交。根据泰泽氏菌的保守序列合成引物扩增泰泽氏菌的基因组DNA,建立了泰泽氏菌的套式PCR方法,并将此方法应用于Scid小鼠和BALB/c小鼠以及可疑样品的检测,其中Scid小鼠的肝、肾组织和BALB/c小鼠的肝组织及可疑组织得到特异性扩增带,其他动物样品和组织DNA均为阴性结果。进一步对该DNA片段进行核酸杂交,结果证实,凡PCR套式扩增为阳性的,均有杂交带。因此,泰泽氏菌套式PCR方法在实际检测中是切实可行,其具有敏感性和特异性高的优点。更多还原

【Abstract】 To apply Nested- PCR on detection of Bacillus piliformis. The Scid and BALB/c mice were intraperitoneally inoculated with RJ strain infected animal tissue. The genomic DNA was amplified by PCR and the fragment of PCR was hybridized. PCR method was established with primers that are conserved among DNA sequences of the Bacillus piliformis. Scid mice、 BALB/c mice and pending samples were monitored with nested PCR, the band was observed in liver,kidney collected from Scid mice and in pending sample, no specific fragments were obtained in other animals. Then, the DNA fragment was hybridized with Dig- labeled probe. Hybridization band was observed in DNA that was positive with PCR. It is possible that Nested PCR can be applied to detect Bacillus piliformis.更多还原