Buchrieser C, Weagant S D, Kaspar C W
Department of Food Microbiology and Toxicology, University of Wisconsin, Madison 53706.
Appl Environ Microbiol. 1994 Dec;60(12):4371-9. doi: 10.1128/aem.60.12.4371-4379.1994.
Sixty strains of Yersinia enterocolitica from five serogroups (O:3; O:9; O:8; O:5; and O:5,27) and eight non-Y. enterocolitica strains, recovered from diverse sources (humans, animals, food, and the environment) in Europe, Argentina, and the United States, were examined by the pulsed-field gel electrophoresis (PFGE) technique of contour clamped homogeneous electric field electrophoresis (CHEF) by using NotI and XbaI as restriction enzymes. NotI and XbaI generated 36 and 33 restriction endonuclease digestion profiles (REDP), respectively. By combining the results of both enzymes, 42 unique genomic groups were differentiated. DNA fragments were transferred to nylon membranes and hybridized with digoxigenin-labelled oligonucleotide probes to the ail gene and virulence plasmid to determine hybridization patterns and the potential virulence of the strains. The strains were tested for the presence of the plasmid by PFGE-CHEF and phenotypic characteristics encoded for by the virulence plasmid. Thirty of the 60 Y. enterocolitica strains tested harbored the virulence plasmid. The specificity of the ail and pYV probes was 100% when tested with 68 Yersinia strains and 19 different non-Yersinia strains. Sixteen selected Y. enterocolitica strains were tested for their virulence by lethality in iron- and desferrioxamine-sensitized mice. No correlation between REDP and the virulence of the strains was observed. The observed REDP and the hybridization patterns were very homogeneous within a serogroup and independent of the source of isolation. In addition, PFGE-CHEF was shown to be valuable in identifying and confirming serogroups. Principal component analysis of Dice similarity indices from REDP was an excellent tool for determining genetic relatedness among strains.
从欧洲、阿根廷和美国不同来源（人类、动物、食品和环境）分离出60株来自5个血清群（O:3、O:9、O:8、O:5和O:5,27）的小肠结肠炎耶尔森菌菌株以及8株非小肠结肠炎耶尔森菌菌株，采用轮廓夹钳均匀电场电泳（CHEF）的脉冲场凝胶电泳（PFGE）技术，以NotI和XbaI作为限制性内切酶进行检测。NotI和XbaI分别产生了36个和33个限制性内切酶消化图谱（REDP）。通过合并两种酶的结果，区分出42个独特的基因组群。将DNA片段转移到尼龙膜上，并用地高辛标记的针对ail基因和毒力质粒的寡核苷酸探针进行杂交，以确定杂交模式和菌株的潜在毒力。通过PFGE-CHEF检测菌株中质粒的存在情况以及毒力质粒编码的表型特征。在检测的60株小肠结肠炎耶尔森菌菌株中，有30株携带毒力质粒。当用68株耶尔森菌菌株和19种不同的非耶尔森菌菌株进行检测时，ail和pYV探针的特异性为100%。通过对铁和去铁胺敏感小鼠的致死率检测16株选定的小肠结肠炎耶尔森菌菌株的毒力。未观察到REDP与菌株毒力之间的相关性。在一个血清群内观察到的REDP和杂交模式非常一致，且与分离来源无关。此外，PFGE-CHEF在鉴定和确认血清群方面很有价值。对REDP的Dice相似性指数进行主成分分析是确定菌株间遗传相关性的极佳工具。
