Tutenel A V, Pierard D, Vandekerchove D, Van Hoof J, De Zutter L
Department of Veterinary Food Inspection, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Vet Microbiol. 2003 Jul 30;94(4):341-6. doi: 10.1016/s0378-1135(03)00121-4.
At present, no standard protocol has been described to detect the presence of Escherichia coli O157 in cattle faeces. Therefore, the sensitivity of 26 different isolation methods was determined in order to recommend a method of choice. Faeces samples from 17 different beef cattle at a farm previously found positive for E. coli O157 were subdivided into a total of 40 samples. It was not known whether the 17 cattle shed E. coli O157 at the time of sampling. At another farm where cattle have been found negative for E. coli O157 on different occasions, five faeces samples were collected. Two methods yielded the highest sensitivity (74%): 6h enrichment in modified tryptone soya broth supplemented with novobiocin (mTSBn) followed by an immunomagnetic separation (IMS) with (i) Dynal beads or (ii) Captivate beads and selective plating on Rainbow agar (RA) plates. Enrichment for 6h was significantly better than 24h enrichment. Only after 24h, buffered peptone water (BPw) was significantly better than mTSBn. A sensitivity of 82% was obtained only when the two most sensitive tests were done simultaneously. Because none of the tests gave 100% sensitivity, it can be concluded that isolation rates of E. coli O157 from bovine faeces using only one of the tested procedures results in an underestimation of the incidence of E. coli O157 in cattle. Performing more than one test on the samples must be considered.
目前,尚未有用于检测牛粪中大肠杆菌O157存在的标准方案。因此,为了推荐一种首选方法,测定了26种不同分离方法的灵敏度。从一个先前检测出大肠杆菌O157呈阳性的农场中选取17头不同肉牛的粪便样本,共分成40个样本。在采样时,并不清楚这17头牛是否携带大肠杆菌O157。在另一个不同时间均检测出牛群大肠杆菌O157呈阴性的农场,采集了5份粪便样本。有两种方法具有最高灵敏度(74%):在添加新生霉素的改良胰蛋白胨大豆肉汤(mTSBn)中富集6小时,然后用(i)Dynal磁珠或(ii)Captivate磁珠进行免疫磁珠分离(IMS),并在彩虹琼脂(RA)平板上进行选择性培养。6小时的富集效果明显优于24小时的富集效果。仅在24小时后,缓冲蛋白胨水(BPw)的效果才明显优于mTSBn。只有同时进行两项最灵敏的检测时,才能获得82%的灵敏度。由于没有一项检测的灵敏度达到100%,因此可以得出结论,仅使用一种测试方法从牛粪中分离大肠杆菌O157会导致对牛群中大肠杆菌O157发病率的低估。必须考虑对样本进行多项检测。