Laboratory of Milk Hygiene and Technology, Department of Food Hygiene and Technology, Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Int J Food Microbiol. 2011 Dec 2;151(2):252-6. doi: 10.1016/j.ijfoodmicro.2011.09.007. Epub 2011 Sep 17.
The transmission pathways of Helicobacter pylori in humans have not been fully elucidated. Research in the last decade has proposed that foodborne transmission, among others, may be a plausible route of human infection. Owing to the organism's fastidious growth characteristics and its ability to convert to viable, yet unculturable states upon exposure to stress conditions, the detection of H. pylori in foods via culture-dependent methods has been proven to be laborious, difficult and in most cases unsuccessful. Hence, nucleic acid-based methods have been proposed as alternative methods but, to date, only PCR-based methods have been reported in the literature. In the current study, fluorescence in situ hybridization (FISH) was used for the detection of H. pylori in raw, bulk-tank bovine milk. After repeated milk centrifugation and washing steps, the bacterial flora of raw milk was subjected to fixation and permeabilization and H. pylori detection was conducted by FISH after hybridization with an H. pylori-specific 16S rRNA-directed fluorescent oligonucleotide probe. Using this protocol, H. pylori was detected in four out of the twenty (20%) raw milk samples examined. The data presented in this manuscript indicate that FISH can serve as an alternative molecular method for screening raw bovine milk for the presence of H. pylori.
幽门螺杆菌在人类中的传播途径尚未完全阐明。过去十年的研究提出，食源性传播可能是人类感染的一种合理途径。由于该生物体生长条件苛刻，并且在暴露于应激条件下能够转化为有活力但无法培养的状态，因此通过依赖培养的方法检测食品中的幽门螺杆菌已被证明是费力、困难且在大多数情况下不成功。因此，已提出基于核酸的方法作为替代方法，但迄今为止，文献中仅报道了基于 PCR 的方法。在本研究中，荧光原位杂交 (FISH) 用于检测生牛乳和散装牛奶奶中的幽门螺杆菌。在重复进行牛奶离心和洗涤步骤后，将生牛乳的细菌菌群进行固定和渗透处理，然后通过与幽门螺杆菌特异性 16S rRNA 导向荧光寡核苷酸探针杂交进行 FISH 检测。使用该方案，在检测的二十（20%）份生牛乳样本中，有四份样本检测到了幽门螺杆菌。本研究中提供的数据表明，FISH 可以作为一种替代分子方法，用于筛选生牛乳中是否存在幽门螺杆菌。
