Use of fluorescent in situ hybridisation for the visualisation of Helicobacter pylori in real drinking water biofilms
Use of fluorescent in situ hybridisation for the visualisation of Helicobacter pylori in real drinking water biofilms
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DOI：
10.2166/wst.2007.282
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发表时间：
2007-01-01
影响因子：
2.7
通讯作者：
Vieira, M. J.
中科院分区：
文献类型：
--
作者：
Braganca, S. M.;Azevedo, N. F.;Vieira, M. J.
荧光标记肽核酸（PNA）探针已被应用于。饮用水生物膜中幽门螺杆菌的原位检测该方法最初应用于从饮用水分配系统（DWDS）中取出的真实的管道样品，但管道的曲率和严重的污垢阻止了通过落射荧光显微镜对细菌的准确检测。因此，将两个半圆形流动池放置在DWDS的旁路中，并定期对暴露长达72天的试样进行采样并分析是否存在H。幽门。在试样的平坦表面中，可以稀疏地检测到表现出与H相似形态的细胞。pylori，其发射PNA探针荧光信号。在杂交程序之前，还在显微镜下观察试样，以用作阴性对照并确保方法的有效性。这项工作证实了已经在其他地方发表的发现，即这种细菌可能存在于DWDS生物膜中。然而，该方法需要训练有素的人员来准确检测病原体，并且在标准水分析实验室中常规使用之前需要简化。
A fluorescently labelled peptide nucleic acid (PNA) probe has been applied for the. in situ detection of Helicobacter pylori in drinking water biofilms. The method was originally applied to real pipe samples removed from a drinking water distribution system (DWDS) but the curvature and the heavy fouling of the pipes prevented an accurate detection of the bacterium by epifluorescence microscopy. Therefore, two semi-circular flow cells were placed in a bypass of the DWDS, and coupons with up to 72 days of exposure were regularly sampled and analysed for the presence of H. pylori. In the flat surfaces of the coupons, it was possible to sparsely detect cells exhibiting similar morphology to H. pylori that were emitting the PNA probe fluorescent signal. Coupons were also visualised under the microscope before the hybridisation procedure to serve as negative controls and ensure the validity of the method. This work corroborates the findings already published elsewhere that this bacterium might be present in DWDS biofilms. The method requires, however, highly trained personnel for an accurate detection of the pathogen and will need simplification before being routinely used in standard water analysis laboratories.