Knutton S, Phillips A D, Smith H R, Gross R J, Shaw R, Watson P, Price E
Institute of Child Health, University of Birmingham, England.
Infect Immun. 1991 Jan;59(1):365-71. doi: 10.1128/iai.59.1.365-371.1991.
The attaching effacing (AE) adherence property is now recognized as an important virulence characteristic of enteropathogenic Escherichia coli (EPEC). The fluorescent-actin staining (FAS) test (S. Knutton, T. Baldwin, P. H. Williams, and A. S. McNeish, Infect. Immun. 57:1290-1298, 1989), which is diagnostic for the AE lesions produced by EPEC (and Vero cytotoxin-producing E. coli), has provided an additional tool with which to investigate this important class of enteric pathogens. In this study, we screened for the AE adherence property in two groups of E. coli isolated from infants with diarrhea by using the FAS test and compared the results with those from O:H serotyping, localized adhesion to HEp-2 cells (LA), and the EPEC adherence factor (EAF) probe. Only 16 of 41 (39%) E. coli strains previously diagnosed as EPEC by O antigen serogrouping were FAS test positive, and of these only 12 belonged to recognized EPEC O:H serotypes; 9 strains which did belong to EPEC O:H serotypes were FAS test negative. Of a second group of 297 untyped E. coli, 7 (2.3%) were FAS test positive, and of these only 2 belonged to EPEC serogroups; 5 belonged to serogroups not regarded as EPEC serogroups or were nontypeable. Of the 23 FAS-test-positive strains identified, 10 were EAF probe positive and showed good LA; 13 were EAF probe negative and showed a quantitatively distinctly poor LA. EAF-positive and EAF-negative strains, however, showed equally good adhesion to human small intestinal mucosa. None of the FAS-test-positive E. coli hybridized with probes for Vero toxins 1 or 2. We conclude that the FAS test is diagnostic not only for classical EPEC and Vero cytotoxin-producing E. coli but also for EPEC strains which are not currently being diagnosed because they belong to serotypes not generally regarded as EPEC serotypes.
紧密黏附(AE)特性现已被公认为肠致病性大肠杆菌(EPEC)的一种重要毒力特征。荧光肌动蛋白染色(FAS)试验(S. 克努顿、T. 鲍德温、P. H. 威廉姆斯和A. S. 麦克尼什,《感染与免疫》57:1290 - 1298,1989年)可用于诊断由EPEC(以及产志贺毒素大肠杆菌)产生的AE损伤,为研究这类重要的肠道病原体提供了又一工具。在本研究中,我们通过FAS试验筛选了两组从腹泻婴儿中分离出的大肠杆菌的AE黏附特性,并将结果与O:H血清分型、对HEp - 2细胞的局部黏附(LA)以及EPEC黏附因子(EAF)探针检测的结果进行了比较。在之前通过O抗原血清分组被诊断为EPEC的41株大肠杆菌中,只有16株(39%)FAS试验呈阳性,其中只有12株属于公认的EPEC O:H血清型;9株属于EPEC O:H血清型的菌株FAS试验呈阴性。在另一组297株未分型的大肠杆菌中,7株(2.3%)FAS试验呈阳性,其中只有2株属于EPEC血清群;5株属于不被视为EPEC血清群的血清群或无法分型。在鉴定出的23株FAS试验阳性菌株中,10株EAF探针呈阳性且LA良好;13株EAF探针呈阴性且LA在数量上明显较差。然而,EAF阳性和EAF阴性菌株对人小肠黏膜的黏附同样良好。FAS试验阳性的大肠杆菌均未与志贺毒素1或2的探针杂交。我们得出结论,FAS试验不仅可用于诊断经典EPEC和产志贺毒素大肠杆菌,还可用于诊断那些因属于一般不被视为EPEC血清型的血清型而目前未被诊断出的EPEC菌株。