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Real-Time PCR Detection of Norovirus in Mussels Collected from the Bosphorus in Istanbul,Turkey
Authors:Huseyin Yilmaz  Kamil Bostan  Nuri Turan  Karlo Muratoglu  Aysun Yilmaz  Ayşe A. Ozkul  Bekir Kocazeybek  Christopher Helps
Affiliation:(1) Department of Virology, Veterinary Faculty, University of Istanbul, Avcilar, Istanbul, Turkey;(2) Department of Food Hygiene and Technology, Veterinary Faculty, University of Istanbul, Avcilar, Istanbul, Turkey;(3) Cevre Industrial Analysis Laboratory, Merkez Mahallesi, Ceylan Sokak, No 24, Mart Plaza, Kat 2, Kagıthane, Istanbul, Turkey;(4) Department of Pediatrics, Zubeyde Hanim Training and Research Center, Baskent University, Izmir, Turkey;(5) Department of Microbiology, Cerrahpasa Faculty of Medicine, University of Istanbul, Cerrahpasa, Istanbul, Turkey;(6) Department of Clinical Veterinary Science, University of Bristol, Langford House, Langford, Bristol, UK
Abstract:Norovirus (NoV) is recognised as one of the most common causes of foodborne infections, and shellfish are a well-documented source of this virus. The presence of NoV in shellfish has not previously been investigated in Turkey, and hence the aim of this study was to determine the frequency of human NoV genogroups I and II in mussels collected from the Bosphorus, Istanbul, Turkey. A total of 320 mussels representing 110 samples originating along the Bosphorus coast were collected from fish distributors. RNA was extracted using the RNeasy Mini Kit and real-time RT–PCR performed using primers specific for NoV genogroup I and II. Amongst the 110 samples, 5 (4.5%) were found to be positive for NoV genogroup II by SYBR Green assay; no genogroup I was detected. A positive signal was obtained by SYBR Green for NoV Genogroup II in mussels collected in October, November and December 2008, and February and July 2009. Only four out of five SYBR Green positive samples could be confirmed by the use of a NoV GII probe-based real-time RT–PCR. The average count and SD of Enterobactericaeae, E. coli and sulphide reductase anaerobic bacteria in PCR positive mussels were 3.56 log ± 0.96 log, 2.32 log ± 0.77 log and 1.70 log ± 0.56 log, respectively. This study shows that NoV Genogroup II is present in mussels collected from the Bosphorus, Istanbul, and may constitute a risk to human health.
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