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在孟加拉国达卡,使用高灵敏度实时聚合酶链反应检测方法未能在饮用水和环境水中检测到幽门螺杆菌DNA。

Failure to detect Helicobacter pylori DNA in drinking and environmental water in Dhaka, Bangladesh, using highly sensitive real-time PCR assays.

作者信息

Janzon Anders, Sjöling Asa, Lothigius Asa, Ahmed Dilruba, Qadri Firdausi, Svennerholm Ann-Mari

机构信息

Department of Microbiology and Immunology, Sahlgrenska Academy at the University of Gothenburg, Box 435, SE-405 30 Gothenburg, Sweden.

出版信息

Appl Environ Microbiol. 2009 May;75(10):3039-44. doi: 10.1128/AEM.02779-08. Epub 2009 Mar 20.

Abstract

The main transmission pathway of Helicobacter pylori has not been determined, but several reports have described detection of H. pylori DNA in drinking and environmental water, suggesting that H. pylori may be waterborne. To address this possibility, we developed, tested, and optimized two complementary H. pylori-specific real-time PCR assays for quantification of H. pylori DNA in water. The minimum detection level of the assays including collection procedures and DNA extraction was shown to be approximately 250 H. pylori genomes per water sample. Using our assays, we then analyzed samples of drinking and environmental water (n = 75) and natural water biofilms (n = 21) from a high-endemicity area in Bangladesh. We could not identify H. pylori DNA in any of the samples, even though other pathogenic bacteria have been found previously in the same water samples by using the same methodology. A series of control experiments were performed to ensure that the negative results were not falsely caused by PCR inhibition, nonspecific assays, degradation of template DNA, or low detection sensitivity. Our results suggest that it is unlikely that the predominant transmission route of H. pylori in this area is waterborne.

摘要

幽门螺杆菌的主要传播途径尚未确定,但有几份报告描述了在饮用水和环境水中检测到幽门螺杆菌DNA,这表明幽门螺杆菌可能通过水传播。为了验证这种可能性,我们开发、测试并优化了两种互补的幽门螺杆菌特异性实时PCR检测方法,用于定量水中的幽门螺杆菌DNA。包括样本采集程序和DNA提取在内,这些检测方法的最低检测水平显示为每个水样约250个幽门螺杆菌基因组。然后,我们使用这些检测方法分析了来自孟加拉国一个高流行地区的饮用水和环境水样本(n = 75)以及天然水生物膜样本(n = 21)。尽管之前使用相同方法在相同水样中发现了其他病原菌,但我们在任何样本中都未检测到幽门螺杆菌DNA。我们进行了一系列对照实验,以确保阴性结果不是由PCR抑制、非特异性检测、模板DNA降解或低检测灵敏度导致的假阴性。我们的结果表明,该地区幽门螺杆菌的主要传播途径不太可能是通过水传播。

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