LABAQUA, S.A., Alicante, Spain.
J Appl Microbiol. 2009 Aug;107(2):416-24. doi: 10.1111/j.1365-2672.2009.04219.x. Epub 2009 Mar 16.
A new real-time PCR assay that simultaneously amplifies a 102-bp fragment of the cagE gene from Helicobacter pylori and a new internal positive control containing a specific sequence of the gyrB gene from Aeromonas hydrophila, was developed and validated for the detection of H. pylori in environmental samples.
The specificity, limits of detection and quantification, repeatability, reproducibility, and accuracy of the method were calculated. The resulting values confirmed the applicability of the method for the quantitative detection of H. pylori. The feasibility of the method was also evaluated by testing 13 pyloric antrum-positive biopsies and 69 water samples, including potable (10), surface (19) and wastewater (40) matrices. The results showed that all the biopsies and 3 of the 40 wastewater samples analysed were positive.
This real-time PCR method provides a sensitive, specific, and accurate method for the rapid quantification of H. pylori in environmental samples.
The PCR diagnostic system proposed in this work, provides a suitable tool for the quantitative detection of H. pylori in environmental samples and can be useful for verifying the role of water as a potential route of its transmission.
开发并验证了一种新的实时 PCR 检测方法，该方法可同时扩增来自幽门螺杆菌的 cagE 基因的 102-bp 片段和来自嗜水气单胞菌的 gyrB 基因的特定序列的新内部阳性对照，用于检测环境样本中的幽门螺杆菌。
计算了该方法的特异性、检测限和定量限、重复性、再现性和准确性。所得值证实了该方法适用于幽门螺杆菌的定量检测。通过测试 13 个幽门窦阳性活检和 69 个水样，包括饮用水（10 个）、地表水（19 个）和废水（40 个）基质，评估了该方法的可行性。结果表明，所有活检和分析的 40 个废水中有 3 个呈阳性。
该实时 PCR 方法为环境样本中幽门螺杆菌的快速定量提供了一种敏感、特异和准确的方法。
本工作中提出的 PCR 诊断系统为环境样本中幽门螺杆菌的定量检测提供了一种合适的工具，可用于验证水作为其传播的潜在途径的作用。
