Boxman Ingeborg L A, Tilburg Jeroen J H C, Te Loeke Nathalie A J M, Vennema Harry, Jonker Klaas, de Boer Enne, Koopmans Marion
Food and Consumer Product Safety Authority, Regionale dienst Oost, P.O. Box 202, 7200 AE, Zutphen, The Netherlands.
Int J Food Microbiol. 2006 May 1;108(3):391-6. doi: 10.1016/j.ijfoodmicro.2006.01.002. Epub 2006 Feb 24.
Shellfish from oyster farms in the Netherlands and imported from other European countries were examined for viral contamination. A method that allows sequence matching between noroviruses from human cases and shellfish was used. The samples of shellfish (n = 42) were analyzed using a semi-nested RT-PCR that had been optimized for detection of norovirus in shellfish (SR primer sets). In addition, a different genome region was targeted using a second primer set which is routinely used for diagnosis of norovirus infection in humans (JV12Y/JV13I). To improve the detection limit for this RT-PCR a semi-nested test format was developed (NV primer sets). One of 21 oyster samples (4.8%) from Dutch farms was norovirus positive, whereas norovirus was detected in 1 out of 8 oyster samples (12.5%) and 5 out of 13 mussel samples (38.5%) collected directly after importation in the Netherlands. RNA from samples associated with an outbreak of gastro-enteritis in the Netherlands in 2001 was re-analyzed using the NV primer sets. At least one identical sequence (142/142 nt) was found in three fecal and in two oyster samples related to this outbreak. Further surveillance of norovirus by detection and typing of viruses from patients with gastroenteritis and shellfish is warranted to clarify the causes of future outbreaks.
对来自荷兰牡蛎养殖场以及从其他欧洲国家进口的贝类进行了病毒污染检测。采用了一种能实现人类病例中的诺如病毒与贝类中的诺如病毒序列匹配的方法。使用针对贝类中诺如病毒检测进行优化的半巢式逆转录聚合酶链反应（SR引物组）对42份贝类样本进行了分析。此外，使用常用于人类诺如病毒感染诊断的另一引物组（JV12Y/JV13I）靶向不同的基因组区域。为提高该逆转录聚合酶链反应的检测限，开发了一种半巢式检测形式（NV引物组）。来自荷兰养殖场的21份牡蛎样本中有1份（4.8%）诺如病毒呈阳性，而在荷兰进口后直接采集的8份牡蛎样本中有1份（12.5%）以及13份贻贝样本中有5份（38.5%）检测到诺如病毒。使用NV引物组对与2001年荷兰一起肠胃炎暴发相关的样本RNA进行了重新分析。在与此暴发相关的3份粪便样本和2份牡蛎样本中发现了至少一个相同序列（142/142核苷酸）。有必要通过对肠胃炎患者和贝类中的病毒进行检测和分型来进一步监测诺如病毒，以明确未来暴发的原因。
