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Applied and Environmental Microbiology, July 2007, p. 4152-4161, Vol. 73, No. 13
0099-2240/07/$08.00+0     doi:10.1128/AEM.00222-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Molecular Identification and Genetic Analysis of Norovirus Genogroups I and II in Water Environments: Comparative Analysis of Different Reverse Transcription-PCR Assays{triangledown}

G. La Rosa,* S. Fontana, A. Di Grazia, M. Iaconelli, M. Pourshaban, and M. Muscillo

Istituto Superiore di Sanità, Department of Environment and Primary Prevention, Roma, Italy

Received 29 January 2007/ Accepted 26 April 2007

Noroviruses have received increased attention in recent years because their role as etiologic agents in acute gastroenteritis outbreaks is now clearly established. Our inability to grow them in cell culture and the lack of an animal model hinder the characterization of these viruses. More recently, molecular approaches have been used to study the genetic relationships that exist among them. In the present study, environmental samples from seawater, estuarine water, and effluents of sewage treatment plants were analyzed in order to evaluate the role of environmental surface contamination as a possible vehicle for transmission of norovirus genogroups I and II. Novel broad-range reverse transcription-PCR/nested assays targeting the region coding for the RNA-dependent RNA polymerase were developed, amplifying fragments of 516 bp and 687 bp in the nested reactions for genogroups II and I, respectively. The assays were evaluated and compared against widely used published assays. The newly designed assays provide long regions for high-confidence BLAST searches in public databases and therefore are useful diagnostic tools for molecular diagnosis and typing of human noroviruses in clinical and environmental samples, as well as for the study of molecular epidemiology and the evolution of these viruses.

* Corresponding author. Mailing address: Istituto Superiore di Sanità, Environmental and Primary Prevention Department, Viale Regina Elena 299, 00161 Rome, Italy. Phone: 39-06-49902718. Fax: 39-06-49902718. E-mail: giuseppina.larosa{at}iss.it

{triangledown} Published ahead of print on 4 May 2007.

Applied and Environmental Microbiology, July 2007, p. 4152-4161, Vol. 73, No. 13
0099-2240/07/$08.00+0     doi:10.1128/AEM.00222-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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