Hennechart-Collette Catherine, Dehan Océane, Fraisse Audrey, Martin-Latil Sandra, Perelle Sylvie
Laboratory for Food Safety, Université Paris-Est, ANSES, 14 rue Pierre et Marie Curie, CEDEX, F-94701 Maisons-Alfort, France.
Microorganisms. 2023 Feb 28;11(3):624. doi: 10.3390/microorganisms11030624.
Viruses are a leading cause of foodborne disease worldwide. Hepatitis viruses (hepatitis A (HAV) and hepatitis E (HEV)) and human norovirus are recognized as the main viruses of public health concern in food hygiene. ISO 15216 approved procedures are not validated for detection of HAV and human norovirus in foodstuffs, such as fishes, leading to an inability to ensure the safety of these products. This study aimed to provide a rapid and sensitive method for detecting these targets in fish products. An existing method that includes proteinase K treatment was selected for further validation using artificially contaminated fish products, according to the recent international standard ISO 16140-4. Recovery efficiencies in pure RNA extracts of viruses ranged from 0.2% to 66.2% for HAV, 4.0% to 100.0% for HEV, 2.2% to 100.0% for norovirus GI, and 0.2% to 12.5% for norovirus GII. LOD values were between 144 and 8.4 × 10 genome copies/g for HAV and HEV, and 10 and 2.0 × 10 copies/g for norovirus GI and norovirus GII, respectively. LOD values were between 3.2 × 10 and 3.6 × 10 genome copies/g for HAV and HEV, and between 8.8 × 10 and 4.4 × 10 genome copies/g for norovirus GI and norovirus GII, respectively. The method developed here was successfully validated in various fish products and can be applied for routine diagnostic needs.
病毒是全球食源性疾病的主要病因。肝炎病毒（甲型肝炎病毒（HAV）和戊型肝炎病毒（HEV））以及人诺如病毒被认为是食品卫生领域主要的公共卫生关注病毒。ISO 15216认可的程序未经验证用于检测食品（如鱼类）中的HAV和人诺如病毒，导致无法确保这些产品的安全性。本研究旨在提供一种快速、灵敏的方法来检测鱼制品中的这些目标病毒。根据最新的国际标准ISO 16140 - 4，选择一种包括蛋白酶K处理的现有方法，使用人工污染的鱼制品进行进一步验证。病毒纯RNA提取物中的回收率，HAV为0.2%至66.2%，HEV为4.0%至100.0%，诺如病毒GI为2.2%至100.0%，诺如病毒GII为0.2%至12.5%。HAV和HEV的检测限（LOD）值分别在144至8.4×10个基因组拷贝/克之间，诺如病毒GI和诺如病毒GII的检测限分别为10至2.0×10个拷贝/克。HAV和HEV的检测限分别在3.2×10至3.6×10个基因组拷贝/克之间，诺如病毒GI和诺如病毒GII的检测限分别在8.8×10至4.4×10个基因组拷贝/克之间。这里开发的方法在各种鱼制品中成功得到验证，可用于常规诊断需求。
