Recovery of structurally intact norovirus from food-contact surfaces

Human noroviruses (NoVs) are the most common cause of gastroenteritis and are responsible for at least 50% of all gastroenteritis outbreaks worldwide. NoV can be transmitted directly via person-to-person contact and contaminated fomites or indirectly via contaminated raw and ready-to-eat foods. However, there have not been enough studies that examine the detection and persistence of NoV on various food-contact surfaces, which may provide information regarding the transmission of NoV in public places, such as restaurants, hospitals, and nursing homes. In order to determine the persistence of NoV, the ability of NoV GII.4 to persist on six kinds of surfaces was investigated for up to 28 d post-inoculation, using an immuno-magnetic separation (IMS) procedure combined with quantitative real-time RT-PCR (qRT-PCR). NoV GII.4 was detected in all test samples, even after 672 h (28 d).     

Transfer of noroviruses between fingers and fomites and food products

Human norovirus (NoV) contaminated hands are important routes for transmission. Quantitative data on transfer during contact with surfaces and food are scarce but necessary for a quantitative risk assessment. Therefore, transfer of MNV1 and human NoVs GI.4 and GII.4 was studied by artificially contaminating human finger pads, followed by pressing on stainless steel and Trespa® surfaces and also on whole tomatoes and cucumber slices. In addition, clean finger pads were pressed on artificially contaminated stainless steel and Trespa® surfaces. The transfers were performed at a pressure of 0.8–1.9 kg/cm² for approximately 2 s up to 7 sequential transfers either to carriers or to food products. MNV1 infectivity transfer from finger pads to stainless steel ranged from 13 ± 16% on the first to 0.003 ± 0.009% on the sixth transfer on immediate transfer. After 10 min of drying, transfer was reduced to 0.1 ± 0.2% on the first transfer to 0.013 ± 0.023% on the fifth transfer. MNV1 infectivity transfer from stainless steel and Trespa® to finger pads after 40 min of drying was 2.0 ± 2.0% and 4.0 ± 5.0% respectively. MNV1 infectivity was transferred 7 ± 8% to cucumber slices and 0.3 ± 0.5% to tomatoes after 10 min of drying, where the higher transfer to cucumber was probably due to the higher moisture content of the cucumber slices. Similar results were found for NoVs GI.4 and GII.4 transfers measured in PCR units. The results indicate that transfer of the virus is possible even after the virus is dried on the surface of hands or carriers. Furthermore, the role of fingers in transmission of NoVs was quantified and these data can be useful in risk assessment models and to establish target levels for efficacy of transmission intervention methods.     

Inactivation of Norovirus Surrogates after Exposure to Atmospheric Ozone

Human norovirus is the leading cause of epidemic gastroenteritis, especially in semiclosed settings such as daycares, nursing homes, hospitals, schools and on cruise ships. Outbreaks are often accompanied by contamination of environmental surfaces and commonly handled items. Surface disinfection of norovirus surrogates, feline calicivirus and murine norovirus, by 20 parts per million atmospheric ozone in a chamber maintaining 80% relative humidity was investigated. After treatment, neither virus could be detected on glass surfaces, but >5 log infectious virus was recovered from untreated controls. Ozone chambers used to decontaminate small, hand-contact items could be an important tool for controlling norovirus outbreaks.     

Detection and molecular characterization of noroviruses from five sewage treatment plants in central Italy

Noroviruses (NoVs) are the most frequent etiological agents of non-bacterial gastroenteritis. These viruses are transmitted through the fecal-oral route, leading to high viral loads in sewages. The objective of this paper was to study the environmental occurrence of the most prevalent NoV strains in different wastewater treatment plants. In addition, molecular characterization of the isolated strains was performed. Two different PCR-based methods were carried out and a novel strategy was used to verify the level of RT-PCR inhibition.From May to September 2007, a total of 97 inflow and outflow samples were collected from five wastewater treatment plants in central Italy. We detected NoV by nested PCR in 96.9% of influent samples: 89.1% contained both genogroups; 4.7% contained only GI and 3.1% only GII. In effluents, we detected NoV in 78.8% of samples: 30.3% contained both genogroups, and 48.5% contained only GI. The major genotypes detected by sequencing analyses were GI/2, GI/5, GII/b, GII/4 and GII/6.This work confirms the wide circulation of NoVs in Italy with a predominance of GI strains, and the widespread distribution of NoV variants in both raw and treated wastewater.     

Investigation of norovirus occurrence in groundwater in metropolitan Seoul, Korea

Groundwater is an important source of drinking and household water worldwide. Hence, the quality of groundwater is very important for preventing waterborne disease outbreaks and should be properly monitored. This study investigated the prevalence of waterborne viruses and fecal indicators in groundwater in metropolitan Seoul and Gyeonggi province, South Korea. A total of 116 samples of groundwater were taken using NanoCeram filters during both summer (June to August) and fall-winter seasons (October to December) in 2008. Among 71 sampling sites, 28 (48.3%) and 18 (35.3%) were positive for norovirus (NoV) from the summer and fall-winter season, respectively. The identified genotypes of NoV include GI-1, 4, 8, 9 and GII-4, 10, 11 (or 17), 13, 15 (or 16). None of fecal indicators was significantly correlated with NoV in groundwater. Among the tested fecal indicators, somatic coliphage (95.3%) showed an excellent true-negative rate of NoV occurrence. The combination of chemical, microbial and viral indicators increased the positive predictive value (50-100%). This study demonstrated a high prevalence of NoV in groundwater in metropolitan Seoul areas and characterized the positive and negative predictive values of a fecal indicator for predicting NoV prevalence.     

Review: norovirus prevalence in Belgian, Canadian and French fresh produce: a threat to human health?

Foodborne viruses, especially noroviruses (NoV), are increasingly reported as the cause of foodborne outbreaks. NoV outbreaks have been reported linked to fresh soft red fruits and leafy greens. Belgium, Canada and France were the first countries to provide data about the prevalence of NoV on fresh produce. In total, 867 samples of leafy greens, 180 samples of fresh soft red fruits and 57 samples of other types of fresh produce (tomatoes, cucumber and fruit salads) were analyzed. Firstly, the NoV detection methodology, including virus and RNA extraction, real-time RT-PCR and quality controls were compared among the three countries. In addition, confirmation and genotyping of the NoV strains was attempted for a subset of NoV positive samples using conventional RT-PCR targeting an alternative region followed by sequencing. Analysis of the process control showed that 653, 179 and 18 samples of the leafy greens, soft red fruits and other fresh produce types were valid for analysis based on the recovery of the process control. NoV was detected by real-time RT-PCR in 28.2% (N = 641), 33.3% (N = 6) and 50% (N = 6) of leafy greens tested in Canada, Belgium and France, respectively. Soft red fruits were found positive by real-time RT-PCR in 34.5% (N = 29) and 6.7% (N = 150) of the samples tested in Belgium and France, respectively. 55.5% (N = 18) of the other fresh produce types, analyzed in Belgium, were found NoV positive by real-time RT-PCR. Conventional RT-PCR resulted in an amplicon of the expected size in 19.5% (52/266) of the NoV positive samples where this assay was attempted. Subsequent sequencing was only successful in 34.6% (18/52) of the suspected amplicons obtained by conventional RT-PCR. From this study, using the described methodology, NoV genomes were frequently detected in fresh produce however sequence confirmation was not successful for the majority of the samples tested. Infection or outbreaks were rarely or not known to be related to the NoV positive samples. With the increase in sensitivity of the detection methodology, there is an increasing concern about the interpretation of positive NoV results by real-time amplification. Strategies to confirm the results by real-time RT-PCR should be developed in analogy with the detection of microbial pathogens in foods. Detection might indicate contact with NoV in the fresh produce chain. Consequently, a potential risk for infection cannot be excluded but the actual risk from RT-PCR NoV positive produce is still unknown. Studies should be designed determining the probability of infection related to the presence or levels of NoV genomic copies.     

2014—2016年济南市哨点医院食源性诺如病毒感染病例流行病学特征分析

目的 了解济南市哨点医院食源性散发腹泻病例中诺如病毒感染类型以及流行病学状况和临床特征。方法 采集2014年1月—2016年12月济南市2家食源性疾病主动监测哨点医院的腹泻病例粪便标本,并收集病例的临床症状资料和流行病学资料。结果 共收集1 292份病例标本,诺如病毒阳性率为18.58%(240/1 292),其中GII基因型占75.42%(181/240)。不同性别的诺如病毒感染病例以及GI和GII基因型病例阳性率差异均无统计学意义(P＞0.05);2种基因型各年龄段阳性率差异有统计学意义(χ²=27.912,P<0.001;χ²=42.285,P<0.001)。第一和第四季度诺如病毒阳性率高于其他季度,具有季节性分布特点。240例诺如病毒阳性病例中以单纯腹泻症状为主(54.17%,130/240),其次为腹泻+呕吐症状(18.75%,45/240);腹泻强度以中度腹泻为主(38.75%,93/240),平均腹泻频次为5.82次/d,粪便性状均以水样便为主(93.75%,225/240)。诺如病毒阳性病例出现呕吐症状的比例为30.00%(72/240),呕吐频次以1～2次/d居多。可疑暴露食物种类以肉与肉制品所占比例最高,其次为水产动物及其制品和乳及乳制品,占比分别为20.00%(48/240)、18.33%(44/240)和15.00%(36/240)。结论 济南市食源性散发腹泻病例诺如病毒感染类型以GII基因型为主,全年流行且具有明显的季节性,婴幼儿为易感人群。应针对诺如病毒感染的季节性特点,以及暴露人群易感程度的差异和暴露食品种类不同的特点,加强食源性散发腹泻病例的监测、识别和防控。     

北京市市售牡蛎中诺如病毒污染对居民健康影响的初步定量风险评估

目的通过建立针对北京市居民从零售到餐桌的牡蛎-诺如病毒暴露模型,对北京市居民通过市售牡蛎暴露诺如病毒的风险开展初步定量评估。方法利用北京市市售牡蛎中诺如病毒的356条定量检测数据,模拟北京市居民牡蛎消费情景,建立暴露评估模型,结合文献发表的基于暴发数据推导的剂量-反应关系模型,对居民通过生食牡蛎后发生诺如病毒感染的健康风险及其影响因素进行评估。结果 GI型和GII型诺如病毒阳性样品的污染水平分别为2.62×104个病毒拷贝/g(95%CI:3.73×103~1.54×105)和5.02×104个病毒拷贝/g(95%CI:8.13×103~2.52×105)。对血清分泌受体阳性的人群,生食1个可能受GI型和GII型诺如病毒污染的牡蛎的发病风险分别为0.93(95%CI:0.73~0.98)和0.95(95%CI:0.80~0.99);对血清分泌受体阴性的人群,生食1个可能污染GI型和GII型诺如病毒牡蛎的发病风险分别为0.37(95%CI:0.04~0.64)和0.57(95%CI:0.07~0.99)。风险的估计值与诺如病毒阴性样品的赋值相关,相关系数为0.49。不确定性分析显示,现有检测方法的检出限高于半数感染及致病剂量,是评估结果不确定性的主要来源。结论北京市居民通过生食牡蛎发生食源性诺如病毒感染的风险很高;提高病毒拷贝定量检测技术的灵敏度,降低检测方法的检出限水平是降低评估结果不确定性的主要途径。     

秦皇岛市部分腹泻患者粪便样本中诺如病毒监测

目的监测秦皇岛市部分诺如病毒引起病毒性腹泻情况,并对阳性样本进行分型。方法收集哨点医院病腹泻患者粪便标本共200份,采用实时荧光定量PCR方法 (real-timeRT-PCR)检测病毒核酸,Mengo病毒作为过程控制和扩增控制,检测诺如病毒回收率和抑制率。结果 Mengo病毒回收率均1%,PCR抑制率≤2%,符合检测要求。检测出病毒阳性15例,其中诺如病毒GⅠ型2例, GⅡ型13例。结论本次采集的样本中诺如病毒流行型分别为GⅠ和GⅡ型,其中以GⅡ组毒株流行为主,诺如病毒阳性率与性别、年龄无关。