China B, Jacquemin E, Devrin A C, Pirson V, Mainil J
Chaire de bactériologie et de pathologie bactérienne, Faculté de médecine vétérinaire, Université de Liège, Belgium.
Res Microbiol. 1999 Jun;150(5):323-32. doi: 10.1016/s0923-2508(99)80058-8.
Enteropathogenic (EPEC) and enterohaemorrhagic (EHEC) Escherichia coli isolated from cattle were studied by DNA colony hybridization to subtype their intimin-encoding (eae) gene with probes derived from the variable parts of the eae alpha gene of the human EPEC strain E2348/69, the eae gamma gene of the human O157:H7 EHEC strain ATCC43888, and the eae beta gene of the bovine O26:H- EHEC strain 193, whose eae gene was first cloned and sequenced during this work. The EPEC and EHEC had been isolated from diarrhoeic calves (143 EPEC and 48 EHEC) and from healthy animals at the slaughterhouse (10 EPEC and 34 EHEC). The 191 bovine EPEC and EHEC isolated from diseased calves were positive with the Eae beta probe (55 and 27% respectively) and with the Eae gamma probe (9 and 73% respectively), whereas 52 EPEC (36%) were negative with the Eae alpha, Eae beta, and Eae gamma probes. The results were different for the 44 bovine EPEC and EHEC isolated from healthy cattle at slaughterhouses: most tested positive with the Eae gamma probe (80 and 82% respectively) and the remaining (20 and 18% respectively) with the Eae beta probe. Nine O26 human EHEC tested positive with the Eae beta probe and seven O111 with the Eae gamma probe. The bovine and human EPEC and EHEC belonging to these two serogroups gave identical results: the 18 bovine and human O26 isolates tested positive with the Eae beta probe, whereas the 13 O111 isolates were positive with the Eae gamma probe. In contrast, the isolates belonging to other serogroups (O5, O15, O18, O20, and O118) gave more variable results. The eae beta and eae gamma, but not the eae alpha, variants were thus distributed amongst bovine EPEC and EHEC. The eae beta variant seemed to be more frequently associated with the presence of clinical signs in calves, but one third of EPEC from diarrhoeic calves carried an eae gene variant other than the alpha, beta, or gamma variants. In addition, the use of these gene probes did not enable differentiation between bovine and human EHEC belonging to the same O serogroup.
采用DNA菌落杂交技术,利用源自人肠致病性大肠杆菌(EPEC)菌株E2348/69的eaeα基因可变区、人O157:H7肠出血性大肠杆菌(EHEC)菌株ATCC43888的eaeγ基因以及牛O26:H - EHEC菌株193的eaeβ基因(其eae基因在本研究中首次被克隆和测序)的探针,对从牛分离出的肠致病性大肠杆菌(EPEC)和肠出血性大肠杆菌(EHEC)进行研究,以对其编码紧密黏附素的(eae)基因进行亚型分类。这些EPEC和EHEC分离自腹泻犊牛(143株EPEC和48株EHEC)以及屠宰场的健康动物(10株EPEC和34株EHEC)。从患病犊牛分离出的191株牛EPEC和EHEC对Eaeβ探针呈阳性反应(分别为55%和27%),对Eaeγ探针也呈阳性反应(分别为9%和73%),而52株EPEC(36%)对Eaeα、Eaeβ和Eaeγ探针均呈阴性。从屠宰场健康牛分离出的44株牛EPEC和EHEC结果不同:大多数对Eaeγ探针呈阳性反应(分别为80%和82%),其余的(分别为20%和18%)对Eaeβ探针呈阳性反应。9株O26人EHEC对Eaeβ探针呈阳性反应,7株O111对Eaeγ探针呈阳性反应。属于这两个血清群的牛和人EPEC及EHEC结果相同:18株牛和人O26分离株对Eaeβ探针呈阳性反应,而13株O111分离株对Eaeγ探针呈阳性反应。相比之下,属于其他血清群(O5、O15、O18、O20和O118)的分离株结果更具变异性。因此,eaeβ和eaeγ变体(而非eaeα变体)分布于牛EPEC和EHEC中。eaeβ变体似乎更常与犊牛临床症状的出现相关,但腹泻犊牛中有三分之一的EPEC携带的eae基因变体不是α、β或γ变体。此外,使用这些基因探针无法区分属于同一O血清群的牛和人EHEC。