Korolik V, Moorthy L, Coloe P J
Department of Applied Biology and Biotechnology, Royal Melbourne Institute of Technology, Victoria, Australia.
J Clin Microbiol. 1995 May;33(5):1136-40. doi: 10.1128/jcm.33.5.1136-1140.1995.
The chromosomal DNA fragment patterns from a total of 169 Campylobacter jejuni and Campylobacter coli isolates from poultry and humans were analyzed by using DNA restriction endonucleases ClaI and EcoRV. The DNA restriction patterns produced by ClaI and EcoRV consisted of unique DNA fragments of 9 to 9.5 kb and 3.5 kb generated with ClaI and a single unique fragment of 3.0 kb produced by EcoRV. These patterns were obtained with all strains of C. jejuni tested. The DNA restriction patterns were further examined by Southern blot analysis with a previously constructed DNA probe, pMO2005, which is also able to distinguish between C. jejuni and C. coli spp. (5). Two types of patterns were produced by hybridization with the ClaI-cleaved DNA of C. jejuni strains, one of a single 18.5-kb genomic fragment and the other of 14.5- and 4.0-kb fragments. This indicated the presence of an extra ClaI site in this genomic fragment in the strains with the duplex pattern. The Southern blot analysis of 169 C. jejuni and C. coli isolates from poultry and from humans with DNA probe pMO2005 demonstrated that 78% of C. jejuni strains isolated from chickens hybridized with DNA probe pMO2005 with a characteristic 14.5- and 4.0-kb banding pattern and 22% hybridized with a single 18.5-kb fragment, whereas 71% of human isolates hybridized with the single 18.5-kb fragment and only 29% hybridized with 14.5- and 4.0-kb fragments. These findings suggest that only a small proportion of C. jejuni strains that colonize chickens may cause disease in humans.
使用DNA限制性内切酶ClaI和EcoRV分析了来自家禽和人类的总共169株空肠弯曲菌和结肠弯曲菌分离株的染色体DNA片段模式。ClaI和EcoRV产生的DNA限制性模式由ClaI产生的9至9.5 kb和3.5 kb的独特DNA片段以及EcoRV产生的3.0 kb单一独特片段组成。所有测试的空肠弯曲菌菌株都获得了这些模式。用先前构建的DNA探针pMO2005通过Southern印迹分析进一步检查DNA限制性模式,该探针也能够区分空肠弯曲菌和结肠弯曲菌属(5)。与空肠弯曲菌菌株的ClaI切割DNA杂交产生了两种类型的模式,一种是单一的18.5 kb基因组片段,另一种是14.5 kb和4.0 kb片段。这表明在具有双链模式的菌株中该基因组片段中存在额外的ClaI位点。用DNA探针pMO2005对来自家禽和人类的169株空肠弯曲菌和结肠弯曲菌分离株进行的Southern印迹分析表明,从鸡中分离的空肠弯曲菌菌株中有78%与DNA探针pMO2005杂交,具有特征性的14.5 kb和4.0 kb条带模式,22%与单一的18.5 kb片段杂交,而71%的人类分离株与单一的18.5 kb片段杂交,只有29%与14.5 kb和4.0 kb片段杂交。这些发现表明,在家禽中定殖的空肠弯曲菌菌株中只有一小部分可能导致人类疾病。