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Applied and Environmental Microbiology, November 2003, p. 6541-6549, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6541-6549.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Reverse Transcription-PCR Analysis of Bottled and Natural Mineral Waters for the Presence of Noroviruses

Gilbert Thierry Lamothe,1* Thierry Putallaz,2 Han Joosten,2 and Joey D. Marugg2

Product Technology Centre, Nestlé Waters Management and Technology, F-88804 Vittel Cedex, France,1 Nestlé Research Center, Vers-chez-les-Blanc, CH-1000 Lausanne 26, Switzerland2

Received 6 December 2002/ Accepted 17 August 2003

A seminested reverse transcription-PCR method coupled to membrane filtration was optimized to investigate the presence of norovirus (NV) RNA sequences in bottled and natural mineral waters. The recovery of viral particles by filtration varied between 28 and 45%, while the limit of detection of the overall method ranged from 6 to 95 viral particles. The assay was broadly reactive, as shown by the successful detection of 27 different viral strains representing 12 common genotypes of NVs. A total of 718 bottled and natural mineral water samples were investigated, including 640 samples of finished, spring, and line products (mostly 1 to 1.5 liters), collected from 36 different water brands of various types and from diverse geographic origins over a 2-year period. In addition, 78 samples of larger volume (10 and 400 to 500 liters) and environmental swabs were investigated. From the 1,436 analyses that were performed for the detection of NVs belonging to genogroups I and II, 34 samples (2.44%) were presumptively positive by seminested RT-PCR. However, confirmation by DNA sequence analysis revealed that all presumptive positive results were either due to nonspecific amplification or to cross-contamination. In conclusion, these results do not provide any evidence for the presence of NV genome sequences in bottled waters.

* Corresponding author: Mailing address: Nestlé Waters M.T., Product Technology Centre, B.P. 101, F-88804 Vittel Cedex, France. Phone: 33/3 29 08 77 28. Fax: 33/3 29 08 71 32. E-mail: Gilbert.Lamothe{at}waters.nestle.com.

Applied and Environmental Microbiology, November 2003, p. 6541-6549, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6541-6549.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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  • Butot, S., Putallaz, T., Croquet, C., Lamothe, G., Meyer, R., Joosten, H., Sanchez, G. (2007). Attachment of Enteric Viruses to Bottles. Appl. Environ. Microbiol. 73: 5104-5110 [Abstract] [Full Text]  
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