Clancy J, Savage D C
Infect Immun. 1981 Apr;32(1):343-52. doi: 10.1128/iai.32.1.343-352.1981.
Two assays were designed with which isogenic laboratory strains of Escherichia coli K-12 with and without ColV plasmids were compared for their ability to adhere in vitro to mouse intestinal epithelium. In both assays, disks of intestinal tissue were exposed to bacteria. In the first, disks were homogenized and the numbers of viable bacteria adherent to them were estimated from colony counts of plates inoculated with dilutions of the homogenates. In the second, bacteria were labeled with [(14)C]aspartic acid; the number of adherent cells per disk was estimated by liquid scintillation spectrometry. Data from each assay were compared by analysis of variance. In both assays, strains bearing the ColV plasmid adhered in two- to threefold-greater numbers than isogenic strains without the plasmid. These differences were highly significant statistically. A non-colicinogenic strain free of the ColV plasmid was selected by treatment of a ColV strain with sodium dodecyl sulfate. In the radioisotopic assay, the ColV strain associated with the epithelium in significantly greater numbers than the cured derivative. A ColV strain was created by conjugation; in the radioisotopic assay this strain bound to epithelium in significantly greater numbers than the recipient strain without the plasmid. The original ColV strain, when negatively stained and examined by electron microscopy, had pili that adsorbed male-specific bacteriophage, whereas its isogenic variant without ColV did not. Some such properties, coded by the plasmid, may increase the virulence of the bacteria.
设计了两种检测方法,用于比较带有和不带有ColV质粒的大肠杆菌K - 12同基因实验室菌株在体外黏附小鼠肠上皮的能力。在这两种检测方法中,肠组织切片都与细菌接触。在第一种方法中,将切片匀浆,通过对接种匀浆稀释液的平板进行菌落计数来估算黏附在切片上的活菌数量。在第二种方法中,用[¹⁴C]天冬氨酸标记细菌;通过液体闪烁光谱法估算每个切片上的黏附细胞数量。通过方差分析比较每种检测方法的数据。在两种检测方法中,携带ColV质粒的菌株黏附数量比不带该质粒的同基因菌株多两到三倍。这些差异在统计学上具有高度显著性。通过用十二烷基硫酸钠处理ColV菌株,筛选出了一株不含ColV质粒的非产大肠杆菌素菌株。在放射性同位素检测中,ColV菌株与上皮细胞结合的数量明显多于经处理后的衍生物。通过接合产生了一株ColV菌株;在放射性同位素检测中,该菌株与上皮细胞结合的数量明显多于不含质粒的受体菌株。对原始的ColV菌株进行负染色并通过电子显微镜检查时,发现其有能吸附雄性特异性噬菌体的菌毛,而其不含ColV的同基因变体则没有。由质粒编码的一些此类特性可能会增加细菌毒力。