Food Safety and Enteric Pathogens Research Unit, National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames 50010, Iowa, USA.
J Appl Microbiol. 2011 Nov;111(5):1283-94. doi: 10.1111/j.1365-2672.2011.05139.x. Epub 2011 Sep 21.
To develop a new adherence assay, using cattle recto-anal junction squamous epithelial (RSE) cells, for evaluating bacterial adherence to cells of bovine origin.
Proof of concept was demonstrated using the human gastrointestinal pathogen Escherichia coli O157:H7, for which cattle are reservoirs. Adherence assays were conducted using both RSE and HEp-2 cells, in the presence and absence of D+Mannose. E. coli O157 specifically adhered in a type I fimbriae-independent manner to RSE cells in significantly higher numbers and also bound significantly higher numbers of RSE cells than diverse laboratory strains of nonpathogenic E. coli.
The RSE cell adhesion assay output highly reproducible and interpretable results that compared very well with those obtained using the more extensively used HEp-2 cell adherence assay.
The RSE cell adhesion assay provides a convenient means of directly defining and evaluating pathogen factors operating at the bovine recto-anal junction. The RSE cell adhesion assay further has the potential for extrapolation to diverse bacteria, including food-borne pathogens that colonize cattle via adherence to this particular anatomical site.
开发一种新的粘附测定法，使用牛直肠-肛门交界处的鳞状上皮（RSE）细胞，评估细菌对牛源细胞的粘附。
使用人胃肠道病原体大肠杆菌 O157：H7（牛是其储主）进行了概念验证。在存在和不存在 D+甘露糖的情况下，使用 RSE 和 HEp-2 细胞进行了粘附测定。大肠杆菌 O157 以一种不依赖于 I 型菌毛的方式特异性地以更高的数量粘附到 RSE 细胞上，并且与多种非致病性大肠杆菌的实验室菌株相比，还结合了更高数量的 RSE 细胞。
RSE 细胞粘附测定法的输出结果高度可重复且易于解释，与更广泛使用的 HEp-2 细胞粘附测定法获得的结果非常吻合。
RSE 细胞粘附测定法为直接定义和评估在牛直肠-肛门交界处起作用的病原体因素提供了一种方便的方法。RSE 细胞粘附测定法还有望推广到多种细菌，包括通过粘附到这个特定解剖部位而定植于牛的食源性病原体。
