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[新鲜农产品中食源性病原体诺如病毒的污染与检测进展]

[The Advances in the Contamination and Detection of Foodborne Pathogen Noroviruses in Fresh Produce].

作者信息

Xie Yajing, Liu Xianjin

出版信息

Bing Du Xue Bao. 2015 Nov;31(6):685-97.

Abstract

This article reviewed the researches proceeding on the contamination and detection of the foodborne pathogen noroviruses (NoVs) in fresh produce, which involved the NoVs contaminations in fresh produce, the special attachment of NoVs in fresh produce, the NoVs outbreaks associated with fresh produce and the NoVs detection in fresh produce. There had been an increase in reported infectious disease risks associated with the consumptions of fresh produce for recent 30 years. Because the NoVs, as a primary cause of viral gastroenteritis thoughout the world, were highly contagious, had a low infectious dose, and were persistent in the environment. And also the methods for NoVs detection in food had significantly developed over the last 15 years. Currently NoVs were the most common pathogen accounting for 40% of outbreaks associated with fresh produce (i. e., fruits and vegetables). Data from outbreaks investigations verified fresh produce as the high risk food products for NoVs. The fresh produce were typically eaten raw with no thermal processing, can be contaminated at any step during production and processing from faecally polluted water and fertilizers, the poor hygiene practices by food handlers and the cross-contamination. The attachment of NoVs to the fresh produce was due to the physio-chemical factors of virus protein coat, the special attachment to different fresh produce, and the possibility for internalization of NoVs. It might provide answers to why those high risk foods were more frequently implicated (i. e., lettuce and raspberries). According to the data of foodborne NoVs outbreaks which were associated with fresh produce from EU countries and the USA, the outbreaks in EU countries were mainly associated with NoVs contaminated raspberries and lettuce, while in USA which were associated with NoVs contaminated lettuce. Unfortunately, there were no NoVs detection methods for fresh produce or the data of foodborne NoVs outbreaks which were associated with fresh produce in China. That made it difficult to analyze the NoVs contamination situation in China. The heterogeneous distributions of presumably low levels of virus, which presented in contaminated fresh produce, also made it difficult to detect NoVs. To solve this problem, different sampling methods, viral elution methods and RT-qPCR methods were chosen. For example, according to the isoelectric point of NoVs particles, high pH and high ionic strength solution could be used as means for releasing NoVs. For the elution from acidic fruit, the buffer capacity and the virus recovery could be increased by the addition of tris-HCl. When analyzing pectin containing raspberries or strawberries, the viral elution usually incubated with pectinase at neutral pH to avoid from foaming jelly. In this paper, the latest ISO standard for NoV detection in food and the new approaches for NoV detection were also reviewed to provide references for domestic researches. It was necessary to establish and develop domestic methods for NoV detection in fresh produce, especially the different NoV conventional molecular detection methods with corresponding NoV extraction methods, which targeted to the different adsorption characteristics of different fruits and vegetables, in order to strengthen the national food safety monitoring.

摘要

本文综述了新鲜农产品中食源性病原体诺如病毒(NoVs)污染及检测方面的研究进展,内容涉及新鲜农产品中的NoVs污染、NoVs在新鲜农产品上的特殊附着、与新鲜农产品相关的NoVs暴发以及新鲜农产品中NoVs的检测。近30年来,与食用新鲜农产品相关的传染病风险报告有所增加。因为诺如病毒作为全球病毒性肠胃炎的主要病因,具有高度传染性、低感染剂量且在环境中持久存在。此外,过去15年里食品中NoVs的检测方法有了显著发展。目前,NoVs是最常见的病原体,占与新鲜农产品(即水果和蔬菜)相关暴发事件的40%。暴发调查数据证实新鲜农产品是NoVs的高风险食品。新鲜农产品通常生食,未经热加工,在生产和加工的任何环节都可能因粪便污染的水和肥料、食品处理人员不良的卫生习惯以及交叉污染而受到污染。NoVs附着于新鲜农产品是由于病毒蛋白衣壳的物理化学因素、对不同新鲜农产品的特殊附着以及NoVs内化的可能性。这或许可以解释为什么那些高风险食品(如生菜和树莓)更常被牵涉其中。根据欧盟国家和美国与新鲜农产品相关的食源性NoVs暴发数据,欧盟国家的暴发主要与NoVs污染的树莓和生菜有关,而在美国则与NoVs污染的生菜有关。遗憾的是,中国尚无新鲜农产品中NoVs的检测方法,也没有与新鲜农产品相关的食源性NoVs暴发数据。这使得难以分析中国的NoVs污染情况。污染新鲜农产品中可能存在的低水平病毒分布不均,也给NoVs检测带来困难。为解决这一问题,选择了不同的采样方法、病毒洗脱方法和RT-qPCR方法。例如,根据NoVs颗粒的等电点,高pH和高离子强度溶液可作为释放NoVs的手段。对于从酸性水果中洗脱,添加tris-HCl可提高缓冲能力和病毒回收率。分析含果胶的树莓或草莓时,病毒洗脱通常在中性pH下与果胶酶一起孵育,以避免果冻起泡。本文还综述了食品中NoV检测的最新ISO标准和NoV检测的新方法,为国内研究提供参考。有必要建立和开发国内新鲜农产品中NoVs的检测方法,特别是针对不同水果和蔬菜不同吸附特性的不同NoV常规分子检测方法及相应的NoV提取方法,以加强国家食品安全监测。

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