食源性诺如病毒在果蔬农产品中的污染及检测研究

发布时间：2018-09-01 10:57

【摘要】：本文综述了食源性致病微生物-诺如病毒(Norovirus,NoV)在果蔬农产品中的来源和分布、在果蔬农产品中的富集情况、与果蔬农产品相关的食源性NoV暴发情况以及在农产品中的检测方法等内容。近30年来,与农产品相关的食源性NoV暴发数目持续增长。这主要与NoV的传播特性(强传染性、低感染剂量和对环境的较强抗性)以及近15年食品中NoV检测方法的显著发展有关。生鲜农产品作为高风险的食品,通常不经烹饪直接生食,在供应链中不同环节都容易被污染,如人类粪便污染的灌溉用水、肥料、食物处理用水、食物处理者的不卫生操作、交叉污染等。NoV在果蔬农产品中的结合主要与病毒颗粒的理化性质及农产品对NoV的吸附和可能的内在化途径有关。根据欧美国家与果蔬农产品有关的食源性NoV暴发的最新数据统计分析,在欧洲大规模食源性暴发主要由被NoV污染的覆盆子和生菜导致,在美国主要由被NoV污染的生菜导致。在国内,由于尚无农产品中食源性NoV检测的相关研究,缺乏与农产品相关NoV暴发的相关数据,因此无法对国内农产品中NoV的污染情况进行具体分析。农产品中NoV具有含量低且分布不均匀的特点,针对不同样品,需要采用合理的采样方法、病毒核糖核酸(Ribonucleic acid,RNA)提取方法以及实时荧光定量PCR(Real-time quantitative polymerase chain reaction,RT-qPCR)方法对其进行检测。根据NoV外壳蛋白的物化特性,高pH值和高离子强度的洗脱溶液(如碱性甘氨酸缓冲液等)可提高NoV的回收率;对于酸性水果中的病毒洗脱,加入三羟甲基氨基甲烷-盐酸(Tris(hydroxymethyl)aminomethane-HCl,Tris-HCl)可以提高回收率;莓果类中含有的果胶,可在中性条件下用果胶酶去除。本文还论述了食品中食源性NoV检测的国际标准ISO/TS 15216:2013、果蔬农产品中NoV检测方法的汇总以及检测NoV的最新方法(原位杂交RT-qPCR方法),为开展农产品中NoV检测的研究提供部分参考,从而建立和发展农产品中NoV的检测方法,针对不同果蔬产品对NoV的吸附特性,配套相应NoV的提取方法,建立多种农产品中NoV常规分子检测方法,以利于对农产品安全的源头监控。
[Abstract]:In this paper, the origin and distribution of foodborne pathogenic microorganism (Norovirus,NoV) in fruit and vegetable produce and its enrichment in fruit and vegetable agricultural products are reviewed. Outbreak of foodborne NoV related to fruit and vegetable farm products and detection methods in agricultural products. In the last 30 years, the number of foodborne NoV outbreaks related to agricultural products continues to increase. This is mainly related to the transmission characteristics of NoV (strong infectivity, low infection dose and strong resistance to the environment) and the remarkable development of NoV detection methods in food in recent 15 years. Fresh produce, as a high-risk food, is usually eaten raw without cooking, and is easily contaminated in different parts of the supply chain, such as irrigation water contaminated with human excrement, fertilizer, water for food treatment, unhygienic operation by food handlers, The binding of. Nov in fruits and vegetables was mainly related to the physicochemical properties of virus particles, the adsorption of NoV to agricultural products and the possible internalization pathway. According to the latest data of foodborne NoV outbreaks related to fruits and vegetables in Europe and the United States, the large scale outbreaks of foodborne NoV in Europe are mainly caused by raspberry and lettuce contaminated by NoV, and mainly by NoV contaminated lettuce in the United States. In our country, there is no research on the detection of foodborne NoV in agricultural products, and there is no relevant data on the outbreak of agri-related NoV, so it is impossible to analyze the pollution of NoV in agricultural products in China. NoV in agricultural products has the characteristics of low content and uneven distribution. It is necessary to use reasonable sampling method, virus ribonucleic acid (Ribonucleic acid,RNA) extraction method and real-time fluorescence quantitative PCR (Real-time quantitative polymerase chain reaction,RT-qPCR) method to detect NoV in different samples. According to the physicochemical properties of NoV shell protein, high pH value and high ionic strength elution solution (such as alkaline glycine buffer, etc.) can improve the recovery of NoV. The recovery rate could be increased by adding trihydroxymethyl aminomethane-hydrochloric acid (Tris (hydroxymethyl) aminomethane-HCl,Tris-HCl), and the pectin contained in berry fruits could be removed by Pectinase under neutral conditions. This paper also discusses the international standard for the detection of foodborne NoV in food, ISO/TS 15216: 2013, the summary of NoV detection methods in fruit and vegetable products and the latest method for detecting NoV (in situ hybridization RT-qPCR method), which provides some references for the research of NoV detection in agricultural products. In order to establish and develop the detection method of NoV in agricultural products, aiming at the adsorption characteristics of NoV in different fruit and vegetable products, matching the corresponding NoV extraction method, establishing the routine molecular detection method of NoV in many kinds of agricultural products. In order to facilitate the safety of agricultural products at the source of monitoring.
【作者单位】： 江苏省食品质量安全重点实验室省部共建国家重点实验室培育基地农业部农产品质量安全控制技术与标准重点实验室江苏省农业科学院食品质量安全与检测所;
【基金】：国家自然科学基金(31301703) 江苏省自主创新项目(CX(14)5068)
【分类号】：R155.5

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