Azevedo N F, Vieira M J, Keevil C W
Environmental Healthcare Unit, School of Biological Sciences, University of Southampton, Southampton, UK.
Water Sci Technol. 2003;47(5):155-60.
Close association of the pathogen Helicobacter pylori in drinking water biofilms has been suggested. Using a two-stage water model, the survival and development of the pathogen in potable water biofilms was monitored. Filter sterilized tap water was used as the growth medium and the inoculum consisted of a naturally occurring consortium of microorganisms. Biofilms were generated on removable stainless steel coupons that were placed in the second vessel. Novel technology peptide nucleic acid (PNA) molecular probes were used to detect and locate the pathogen in the biofilms. The PNA-labelled oligonucleotide probes were highly specific, and complementary to the helix 6 region of H. pylori 16S rRNA. The pathogen was tracked in the biofilms using epifluorescence microscopy and episcopic differential interference contrast microscopy. Results show that H. pylori can successfully incorporate within biofilms and its presence was detected for up to five days after inoculation. PNA probes provided an easy and quick way of performing fluorescence in situ hybridisation assays in heterogeneous biofilms.
有人提出饮用水生物膜中存在病原体幽门螺杆菌。使用两阶段水模型，监测了该病原体在饮用水生物膜中的存活和生长情况。过滤除菌的自来水用作生长培养基，接种物由天然存在的微生物群落组成。生物膜在放置于第二个容器中的可移动不锈钢试片上形成。采用新技术肽核酸（PNA）分子探针检测和定位生物膜中的病原体。PNA标记的寡核苷酸探针具有高度特异性，与幽门螺杆菌16S rRNA的螺旋6区域互补。使用落射荧光显微镜和落射微分干涉对比显微镜追踪生物膜中的病原体。结果表明，幽门螺杆菌能够成功地融入生物膜中，并且在接种后长达五天都能检测到其存在。PNA探针为在异质生物膜中进行荧光原位杂交分析提供了一种简便快捷的方法。
