Schmidt H, Plaschke B, Franke S, Rüssmann H, Schwarzkopf A, Heesemann J, Karch H
Institut für Hygiene und Mikrobiologie, Universität Würzburg, Germany.
Med Microbiol Immunol. 1994 Feb;183(1):23-31. doi: 10.1007/BF00193628.
In this study 98 Escherichia coli strains which belonged to traditional enteropathogenic (EPEC) serotypes and 82 enterohemorrhagic E. coli (EHEC) strains were screened by polymerase chain reaction (PCR) for the presence of E. coli-attaching and -effacing (eae) genes. These strains were also hybridized with the enteropathogenic adherence factor (EAF) probe and examined in the fluorescence actin staining (FAS) test. The results obtained from the individual strains demonstrated that all 26 class I EPEC with localized adherence to HEp-2 cells carried EAF and eae genes. In contrast, of 72 EPEC strains with no or diffuse adherence only 1 strain was EAF positive and 6 strains had eae. Of 82 EHEC strains a total of 75 carried eae sequences. Of considerable interest, 15 of 21 E. coli strains that lost their slt genes during subcultivation were found to be eae positive. As controls a total of 53 enterotoxigenic and enteroinvasive E. coli, and 125 E. coli strains from the normal flora were investigated and all displayed negative results in the eae-PCR. From the 201 strains comprising classical EPEC serotypes, EHEC and E. coli with lost slt genes, a total of 126 displayed a positive FAS test and 122 reacted in the eae-PCR. Only 4 strains were FAS test positive but eae-PCR negative. Our data indicate that E. coli strains possessing the eae genes are heterogenous with respect to their virulence determinants. Loss of virulence plasmids and phage-encoded slt genes either in the host or during storage may contribute to this heterogeneity. The eae-PCR detected all class I EPEC and 91.5% of the EHEC.(ABSTRACT TRUNCATED AT 250 WORDS)
在本研究中,通过聚合酶链反应(PCR)对98株属于传统肠致病性大肠杆菌(EPEC)血清型的菌株和82株肠出血性大肠杆菌(EHEC)菌株进行筛选,以检测是否存在大肠杆菌黏附和脱落(eae)基因。这些菌株还与肠致病性黏附因子(EAF)探针杂交,并进行荧光肌动蛋白染色(FAS)试验。从各个菌株获得的结果表明,所有26株对HEp-2细胞有局部黏附的I类EPEC均携带EAF和eae基因。相比之下,在72株无黏附或弥散性黏附的EPEC菌株中,只有1株EAF呈阳性,6株有eae基因。在82株EHEC菌株中,共有75株携带eae序列。相当有趣的是,在传代培养过程中失去slt基因的21株大肠杆菌菌株中有15株被发现eae呈阳性。作为对照,共研究了53株产肠毒素和侵袭性大肠杆菌以及125株来自正常菌群的大肠杆菌菌株,所有菌株在eae-PCR中均显示阴性结果。在由经典EPEC血清型、EHEC和失去slt基因的大肠杆菌组成的201株菌株中,共有126株FAS试验呈阳性,122株在eae-PCR中呈阳性反应。只有4株FAS试验呈阳性但eae-PCR呈阴性。我们的数据表明,具有eae基因的大肠杆菌菌株在其毒力决定因素方面是异质的。宿主中或储存期间毒力质粒和噬菌体编码的slt基因的丢失可能导致这种异质性。eae-PCR检测到了所有I类EPEC和91.5%的EHEC。(摘要截短至250字)