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Applied and Environmental Microbiology, May 2009, p. 3039-3044, Vol. 75, No. 10
0099-2240/09/$08.00+0     doi:10.1128/AEM.02779-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Failure To Detect Helicobacter pylori DNA in Drinking and Environmental Water in Dhaka, Bangladesh, Using Highly Sensitive Real-Time PCR Assays{triangledown}

Anders Janzon,1* Åsa Sjöling,1 Åsa Lothigius,1 Dilruba Ahmed,2 Firdausi Qadri,2 and Ann-Mari Svennerholm1

Department of Microbiology and Immunology, Sahlgrenska Academy at the University of Gothenburg, Box 435, SE-405 30 Gothenburg, Sweden,1 International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B), GPO Box 128, Dhaka 1000, Bangladesh2

Received 6 December 2008/ Accepted 13 March 2009

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.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Gothenburg, Box 435, SE-405 30 Gothenburg, Sweden. Phone: 46-31-7866203. Fax: 46-31-7866205. E-mail: anders.janzon{at}microbio.gu.se

{triangledown} Published ahead of print on 20 March 2009.

Applied and Environmental Microbiology, May 2009, p. 3039-3044, Vol. 75, No. 10
0099-2240/09/$08.00+0     doi:10.1128/AEM.02779-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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