【摘要】：本文綜述了食源性致病微生物-諾如病毒(Norovirus,NoV)在果蔬農(nóng)產(chǎn)品中的來(lái)源和分布、在果蔬農(nóng)產(chǎn)品中的富集情況、與果蔬農(nóng)產(chǎn)品相關(guān)的食源性NoV暴發(fā)情況以及在農(nóng)產(chǎn)品中的檢測(cè)方法等內(nèi)容。近30年來(lái),與農(nóng)產(chǎn)品相關(guān)的食源性NoV暴發(fā)數(shù)目持續(xù)增長(zhǎng)。這主要與NoV的傳播特性(強(qiáng)傳染性、低感染劑量和對(duì)環(huán)境的較強(qiáng)抗性)以及近15年食品中NoV檢測(cè)方法的顯著發(fā)展有關(guān)。生鮮農(nóng)產(chǎn)品作為高風(fēng)險(xiǎn)的食品,通常不經(jīng)烹飪直接生食,在供應(yīng)鏈中不同環(huán)節(jié)都容易被污染,如人類糞便污染的灌溉用水、肥料、食物處理用水、食物處理者的不衛(wèi)生操作、交叉污染等。NoV在果蔬農(nóng)產(chǎn)品中的結(jié)合主要與病毒顆粒的理化性質(zhì)及農(nóng)產(chǎn)品對(duì)NoV的吸附和可能的內(nèi)在化途徑有關(guān)。根據(jù)歐美國(guó)家與果蔬農(nóng)產(chǎn)品有關(guān)的食源性NoV暴發(fā)的最新數(shù)據(jù)統(tǒng)計(jì)分析,在歐洲大規(guī)模食源性暴發(fā)主要由被NoV污染的覆盆子和生菜導(dǎo)致,在美國(guó)主要由被NoV污染的生菜導(dǎo)致。在國(guó)內(nèi),由于尚無(wú)農(nóng)產(chǎn)品中食源性NoV檢測(cè)的相關(guān)研究,缺乏與農(nóng)產(chǎn)品相關(guān)NoV暴發(fā)的相關(guān)數(shù)據(jù),因此無(wú)法對(duì)國(guó)內(nèi)農(nóng)產(chǎn)品中NoV的污染情況進(jìn)行具體分析。農(nóng)產(chǎn)品中NoV具有含量低且分布不均勻的特點(diǎn),針對(duì)不同樣品,需要采用合理的采樣方法、病毒核糖核酸(Ribonucleic acid,RNA)提取方法以及實(shí)時(shí)熒光定量PCR(Real-time quantitative polymerase chain reaction,RT-qPCR)方法對(duì)其進(jìn)行檢測(cè)。根據(jù)NoV外殼蛋白的物化特性,高pH值和高離子強(qiáng)度的洗脫溶液(如堿性甘氨酸緩沖液等)可提高NoV的回收率;對(duì)于酸性水果中的病毒洗脫,加入三羥甲基氨基甲烷-鹽酸(Tris(hydroxymethyl)aminomethane-HCl,Tris-HCl)可以提高回收率;莓果類中含有的果膠,可在中性條件下用果膠酶去除。本文還論述了食品中食源性NoV檢測(cè)的國(guó)際標(biāo)準(zhǔn)ISO/TS 15216:2013、果蔬農(nóng)產(chǎn)品中NoV檢測(cè)方法的匯總以及檢測(cè)NoV的最新方法(原位雜交RT-qPCR方法),為開(kāi)展農(nóng)產(chǎn)品中NoV檢測(cè)的研究提供部分參考,從而建立和發(fā)展農(nóng)產(chǎn)品中NoV的檢測(cè)方法,針對(duì)不同果蔬產(chǎn)品對(duì)NoV的吸附特性,配套相應(yīng)NoV的提取方法,建立多種農(nóng)產(chǎn)品中NoV常規(guī)分子檢測(cè)方法,以利于對(duì)農(nóng)產(chǎn)品安全的源頭監(jiān)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.
【作者單位】： 江蘇省食品質(zhì)量安全重點(diǎn)實(shí)驗(yàn)室省部共建國(guó)家重點(diǎn)實(shí)驗(yàn)室培育基地農(nóng)業(yè)部農(nóng)產(chǎn)品質(zhì)量安全控制技術(shù)與標(biāo)準(zhǔn)重點(diǎn)實(shí)驗(yàn)室江蘇省農(nóng)業(yè)科學(xué)院食品質(zhì)量安全與檢測(cè)所;
【基金】：國(guó)家自然科學(xué)基金(31301703) 江蘇省自主創(chuàng)新項(xiàng)目(CX(14)5068)
【分類號(hào)】：R155.5

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