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Applied and Environmental Microbiology, September 2007, p. 5464-5470, Vol. 73, No. 17
0099-2240/07/$08.00+0     doi:10.1128/AEM.00572-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Sensitive Multiplex Real-Time Reverse Transcription-PCR Assay for the Detection of Human and Animal Noroviruses in Clinical and Environmental Samples{triangledown}

Sandro Wolf,1 Wendy M. Williamson,2 Joanne Hewitt,1 Malet Rivera-Aban,1 Susan Lin,2 Andrew Ball,2 Paula Scholes,2 and Gail E. Greening1*

Communicable Disease Group, Institute of Environmental Science & Research Ltd., Kenepuru Science Centre, P.O. Box 50-348, Porirua, New Zealand,1 Water Resource Group, Institute of Environmental Science & Research Ltd., Christchurch Science Centre, P.O. Box 29-181, Christchurch, New Zealand2

Received 13 March 2007/ Accepted 28 June 2007

In this study, we developed a triplex real-time reverse transcription-PCR (RT-PCR)-based method that detects and distinguishs between noroviruses belonging to genogroups I, II, and III and that targets the junction between the regions of open reading frame 1 (ORF1) and ORF2. This is the first assay to include all three genogroups and the first real-time RT-PCR-based method developed for the detection of bovine noroviruses. The assay was shown to be broadly reactive against a wide spectrum of norovirus genotypes, including GI/1 through GI/7, GII/1 through GII/8, GII/10, GII/12, and GII/17, in different matrices (including fecal specimens, treated and raw sewage, source water, and treated drinking water). The assay is highly sensitive, detecting low copy numbers of plasmids that carry the target sequence. A new bovine norovirus, Bo/NLV/Norsewood/2006/NZL, was identified by this assay and was further genetically characterized. The results implicate a broad range of possible applications, including clinical diagnostics, tracing of fecal contaminants, and due to its sensitivity and broad reactivity, environmental studies.

* Corresponding author. Mailing address: Institute of Environmental Science and Research Ltd., Kenepuru Science Centre, P.O. Box 50-348, Porirua, New Zealand. Phone: 64-4-9140765. Fax: 64-4-9140770. E-mail: Gail.greening{at}esr.cri.nz

{triangledown} Published ahead of print on 6 July 2007.

Applied and Environmental Microbiology, September 2007, p. 5464-5470, Vol. 73, No. 17
0099-2240/07/$08.00+0     doi:10.1128/AEM.00572-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hewitt, J., Bell, D., Simmons, G. C., Rivera-Aban, M., Wolf, S., Greening, G. E. (2007). Gastroenteritis Outbreak Caused by Waterborne Norovirus at a New Zealand Ski Resort. Appl. Environ. Microbiol. 73: 7853-7857 [Abstract] [Full Text]  


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