Szabo Kathrin, Trojnar Eva, Anheyer-Behmenburg Helena, Binder Alfred, Schotte Ulrich, Ellerbroek Lüppo, Klein Günter, Johne Reimar
Institute of Food Quality and Food Safety, University of Veterinary Medicine Hannover, Hannover, Germany,; Unit Food Hygiene and Virology, Federal Institute for Risk Assessment, Berlin, Germany.
Unit Food Hygiene and Virology, Federal Institute for Risk Assessment, Berlin, Germany.
Int J Food Microbiol. 2015 Dec 23;215:149-56. doi: 10.1016/j.ijfoodmicro.2015.09.013. Epub 2015 Sep 21.
Hepatitis E virus (HEV) is a pathogen of increasing importance, which can be zoonotically transmitted from domestic pigs, wild boar, and deer to humans. Foodborne transmission by consumption of raw and undercooked liver, meat, or sausages prepared from infected animals has been documented. The aim of this study was to investigate the distribution of HEV in different types of sausages sold in Germany. As no standardized methods for HEV detection in food exist, several techniques of sample homogenization, virus concentration and nucleic acid extraction followed by real-time RT-PCR were compared using artificially contaminated sausages. A method using TRI Reagent® Solution showed the best efficacy of matrix disruption and a treatment with chloroform followed by a silica-based RNA extraction method resulted in the highest HEV detection rates. The detection limit of the method was 2.9 × 10(3) and 5.3 × 10(4) genome equivalents per 5 g raw sausage and 2 g liver sausage, respectively. Application of the method to raw and liver sausages from retail in Germany resulted in the HEV genome detection in 14 out of 70 (20%) raw sausages and in 11 out of 50 (22%) liver sausages. The detected HEV sequences showed a high diversity and belonged to different subtypes of HEV genotype 3. The results indicate a broad distribution of HEV-RNA in meat products sold in Germany; however, the infectivity of the detected virus remains to be assessed in future.
戊型肝炎病毒（HEV）是一种重要性日益增加的病原体，它可通过人畜共患病的方式从家猪、野猪和鹿传播给人类。食用由受感染动物制成的生的和未煮熟的肝脏、肉类或香肠导致的食源性传播已有记录。本研究的目的是调查在德国销售的不同类型香肠中戊型肝炎病毒的分布情况。由于不存在用于食品中戊型肝炎病毒检测的标准化方法，因此使用人工污染的香肠比较了几种样品匀浆、病毒浓缩和核酸提取技术，随后进行实时逆转录聚合酶链反应。使用TRI Reagent®溶液的方法显示出最佳的基质破坏效果，用氯仿处理后再采用基于硅胶的RNA提取方法可获得最高的戊型肝炎病毒检测率。该方法的检测限分别为每5克生香肠2.9×10³个基因组当量和每2克肝香肠5.3×10⁴个基因组当量。将该方法应用于德国零售的生香肠和肝香肠，结果在70份生香肠中有14份（20%）检测到戊型肝炎病毒基因组，在50份肝香肠中有11份（22%）检测到。检测到的戊型肝炎病毒序列显示出高度的多样性，属于戊型肝炎病毒3型的不同亚型。结果表明戊型肝炎病毒RNA在德国销售的肉类产品中广泛分布；然而，检测到的病毒的传染性仍有待在未来进行评估。
