实时荧光PCR检测诺瓦克病毒的应用研究

目的应用实时荧光PCR方法对诺瓦克病毒性急性胃肠炎进行快速检测，为疾病预防控制提供准确可靠的实验诊断依据。方法采集一起急性胃肠炎暴发流行患者的1份粪便、5份肛拭和1份呕吐物，进行实时荧光PCR检测。结果实时荧光PCR检测有1份病人粪便和1份病人肛拭样本中诺瓦克病毒核酸呈阳性。结论这是一起由诺瓦克病毒感染引起的急性胃肠炎暴发流行。     

广东省2005年诺瓦克样病毒感染暴发疫情

目的分析广东省2005年群体性诺瓦克样病毒性胃肠炎流行特征,为今后预防控制提供科学依据。方法采用描述性分析方法,对广东省2005年群体性诺瓦克样病毒性胃肠炎疫情进行调查分析。结果2005年广东省共报告12起群体性诺瓦克样病毒性胃肠炎疫情,共发病469例,无死亡病例。疫情全部发生在学校。发生时间除1起发生在3月份外,11起集中发生在9~12月份。症状以呕吐为主。12起疫情中病例数最多的1起达190例,罹患率最高的1起达53.3%(23/43)。11起疫情患者有共同的进餐史,1起疫情患者没有共同的食物暴露史。每起疫情均从部分患者粪便标本中检出诺瓦克样病毒核酸或抗原。结论学校是诺瓦克样病毒暴发疫情的好发地方,食物传播、接触呕吐物和粪便传播是可能的主要传播途径,因此控制诺瓦克样病毒暴发疫情的关键是抓好学校的饮食卫生和妥善处理呕吐物和粪便。     

群体性诺瓦克样病毒性胃肠炎疫情分析

目的 分析广州市2003年群体性诺瓦克样病毒性胃肠炎流行特征，加强该病的预防控制工作。方法 采用现场流行病学调查方法，对广州市2003年群体性诺瓦克样病毒性胃肠炎疫情进行调查分析。结果 2003年10月～12月底，广州市共报告8起群体性诺瓦克样病毒性胃肠炎疫情，共计病例396例，每起疫情均从部分患者粪便标本中检出诺瓦克病毒核酸或抗原。结论 RT-PCR的阳性检出率大于ELISA法，粪便标本检出率高。诺瓦克样病毒性胃肠炎常引起暴发疫情，及时采取控制措施可避免疫情扩散。     

诺瓦克样病毒胃肠炎集体性爆发的证实与思考

目的通过对诺瓦克样病毒胃肠炎集体爆发的调查和证实,总结处理不明原因疾病的经验.方法运用流行病学方法对3所幼儿园和1所小学发生的急性胃肠炎集体爆发进行调查,对病人粪便样本进行诺瓦克样病毒核酸检验.结果4起集体爆发的胃肠炎均以恶心呕吐为主要表现,有2个发病高峰,病程较短,有自愈倾向,没有明显的年龄、班级、楼层分布的集中趋势.对第4起爆发的22份病人粪便样本进行诺瓦克病毒的PCR检测,其中19份阳性,同时有4份经DNA测序证实与诺瓦克样病毒同源.结论此一系列不明原因的胃肠炎是由诺瓦克样病毒引起的.这是我国证实由诺瓦克样病毒感染而引起的集体性急性胃肠炎的爆发.     

一起诺瓦克样病毒性胃肠炎的流行病学调查

目的探讨2004年广州市番禺区一起由诺瓦克样病毒引起的学校胃肠炎疫情暴发特征. 方法采用现场流行病学调查方法,对该起疫情进行调查分析. 结果该起疫情共报告病例30例 ,其中3例患者的粪便或肛拭标本检出诺瓦克病毒抗原. 结论本次胃肠炎暴发疫情由诺瓦克样病毒引起,传染来源尚未查明.     

广州市1起诺如病毒急性胃肠炎暴发的实验室检测

目的确定1起学校急性胃肠炎暴发原因。方法采集病例粪便标本,直饮水标本,采用实时荧光定量(Real-time)PCR和RT-PCR方法检测病毒核酸,阳性标本测序分析。结果 5份粪便标本中3份呈诺如病毒阳性,2份直饮水标本均呈诺如病毒阳性。3份病例粪便标本病毒株核苷酸序列一致,均为GⅡ-4型,提示病毒同源。结论对诺如病毒进行核酸检测,证实该疫情为一起诺如病毒GⅡ-4型引起的急性胃肠炎暴发。     

一起福利院诺瓦克病毒胃肠炎暴发调查

目的调查分析2006年嘉定区一起由诺瓦克病毒引起的福利院胃肠炎暴发疫情的特征及其原因。方法采用现场流行病学方法进行调查,对标本采用RT-PCR方法检测诺瓦克病毒核酸。结果该起疫情共报告病例53例,52例集中在1号楼,1号楼的罹患率为24.41%,在7份病人标本中3份检出诺瓦克病毒核酸阳性。结论本次胃肠炎暴发疫情是由诺瓦克病毒引起,传播途径为人与人之间密切接触传播。     

一起养老院诺瓦克样病毒胃肠炎暴发的调查

诺瓦克样病毒是急性胃肠炎的主要病原之一,全年均可发病,但冬季较多,一般在学校、家庭、托幼机构、养老院等人群容易出现暴发.2008年1月22日至2月2日广州市一家养老院发生一起诺瓦克样病毒引起的胃肠炎疫情. 1.材料与方法:采用《广东省群体性诺瓦克病毒胃肠炎个案调查表》对发病者进行调查,发病后1～3天取粪便标本,冷藏送广州市疾病预防控制中心病毒室进行RT-PCR检测.     

一起养老院诺瓦克样病毒胃肠炎暴发的调查

诺瓦克样病毒是急性胃肠炎的主要病原之一,全年均可发病,但冬季较多,一般在学校、家庭、托幼机构、养老院等人群容易出现暴发.2008年1月22日至2月2日广州市一家养老院发生一起诺瓦克样病毒引起的胃肠炎疫情. 1.材料与方法:采用《广东省群体性诺瓦克病毒胃肠炎个案调查表》对发病者进行调查,发病后1～3天取粪便标本,冷藏送广州市疾病预防控制中心病毒室进行RT-PCR检测.     

一起养老院诺瓦克样病毒胃肠炎暴发的调查

诺瓦克样病毒是急性胃肠炎的主要病原之一,全年均可发病,但冬季较多,一般在学校、家庭、托幼机构、养老院等人群容易出现暴发.2008年1月22日至2月2日广州市一家养老院发生一起诺瓦克样病毒引起的胃肠炎疫情. 1.材料与方法:采用《广东省群体性诺瓦克病毒胃肠炎个案调查表》对发病者进行调查,发病后1～3天取粪便标本,冷藏送广州市疾病预防控制中心病毒室进行RT-PCR检测.     

一起养老院急性胃肠炎疫情的病原学分析

目的明确一起养老院感染性腹泻疫情的病原体及其基因型别。方法通过流行病学调查,查明罹患率。对发病老人和部分工作人员进行肛拭子采样,采用细菌培养检测常规肠道致病菌,应用胶体金法检测轮状病毒,采用实时荧光定量RT-PCR进行诺如病毒核酸检测,应用常规RT-PCR扩增部分NoV阳性株衣壳蛋白的N/S区,PCR产物纯化回收、测序,利用Clustal X、MEGA4.1软件分析其基因序列。结果 21份病例及工作人员肛拭子样本,肠道致病菌和轮状病毒均为阴性,19份样本诺如病毒核酸阳性,毒株排除重组株的可能,测序样本均属GⅡ.4型诺如病毒,其与Hu/NSW696T/2006/AUS（EF684915）的同源性高达99%。结论经诺如病毒核酸序列分析,该疫情是一起由GⅡ.4-2006b型诺如病毒引起的急性胃肠炎暴发。     

一起邮轮诺如病毒痰隋的病原学检测

[目的]总结诺如病毒实验室诊断经验,进一步完善诺如病毒的检测方法。（方法]针对上海口岸发生的又1起邮轮急性胃肠炎爆发疫情,应用酶联免疫吸附实验（ELISA）、逆转录-聚合酶链反应（RT—PCR）、电镜观察等技术,对送检的9份粪便标本进行检测。[结果]运用2种ELISA试剂盒在9份标本中分别检出4份和5份标本诺如病毒抗原阳性;运用逆转录-聚合酶链方法,发现9份标本中有7份扩增出诺如病毒特异性条带;运用电镜在镜下观察到诺如病毒颗粒。[结论]诺如病毒检测方法的建立将为口岸公共卫生突发事件应急体系的建设提供重要的技术支持。     

Emerging genotype (GGIIb) of norovirus in drinking water, Sweden

From May through June 2001, an outbreak of acute gastroenteritis that affected at least 200 persons occurred in a combined activity camp and conference center in Stockholm County. The source of illness was contaminated drinking water obtained from private wells. The outbreak appears to have started with sewage pipeline problems near the kitchen, which caused overflow of the sewage system and contaminated the environment. While no pathogenic bacteria were found in water or stools specimens, norovirus was detected in 8 of 11 stool specimens and 2 of 3 water samples by polymerase chain reaction. Nucleotide sequencing of amplicons from two patients and two water samples identified an emerging genotype designated GGIIb, which was circulating throughout several European countries during 2000 and 2001. This investigation documents the first waterborne outbreak of viral gastroenteritis in Sweden, where nucleotide sequencing showed a direct link between contaminated water and illness.     

2013年广州市某小学诺如病毒感染痰隋的流行病学分析

目的探讨诺如病毒感染引起的急性胃肠炎的暴发特点和流行原因,为制定防控策略提供依据。方法采用现场流行病学调查方法,结合临床表现及实验室检测对广州市某小学暴发的诺如病毒引起的感染性腹泻疫情进行流行病学分析。结果2013年12月23—26日广州市某小学共发生急性胃肠炎病例107例,均为学生,罹患率9．18％;95．33％（102／107）表现为呕吐,46．73％（50／107）为发热,27．10％为（29／107）腹泻,症状较轻。进行校外托管是该起疫情的危险因素（OR=3．215,95％CI：1．150-8．987,X^2=5．107,P〈0．05）。采集的12例病例粪便／肛拭子样本,7例为诺如病毒GⅡ型阳性。结论在校外托管班进行午餐和午休托管可能是本次诺如病毒感染疫情发生的危险因素,校外托管班应引起教育和卫生部门的关注。     

An outbreak of Salmonella gastroenteritis in an urban jail

An outbreak of gastroenteritis in New York City's largest jail involved 145 cases over a two-month period. The outbreak was unusual in that two Salmonella strains (serogroups B and D) were involved. Management of the outbreak involved screening kitchen workers by culture of stool samples, and education regarding personal hygiene. Obstacles to investigation and management of the outbreak arose out of the special nature of the jail environment; these included jurisdictional problems and high turnover of the inmate population.     

Wading pool water contaminated with both noroviruses and astroviruses as the source of a gastroenteritis outbreak

In July 2001, an outbreak of gastroenteritis occurred in Helsinki among children and adults after bathing in an outdoor wading pool. The epidemiological survey revealed that at least 242 persons were affected. Microbiological testing of both patient stool samples and of the pool water revealed the presence of two different gastroenteritis viruses: a norovirus (NV) and an astrovirus. Amplicon sequencing of the NV samples showed nucleotide sequence identity between the virus from patients and the water. After changing the pool water and the sand at the bottom of the pool followed by shock chlorination, no virus could be detected in the water. However, NV was continuously detected in the water outlet well as much as 8 months after the incident. Here we show how molecular methods aided in tracing the source of the epidemic and in finding the causative pathogens both in patients and in the environment.     

诺如病毒RdRp区和核心区部分基因克隆及序列分析

目的:明确引起急性胃肠炎暴发的诺如病毒RdRp区和衣壳区部分核酸序列,确定病毒基因亚型。方法:采集腹泻病人粪便标本,提取病毒核酸,通过RT-PCR扩增长度为387 bp的病毒RNA多聚酶区(RdRp)区和核心区部分核酸序列,并克隆至T载体,转化大肠杆菌,筛选阳性菌落,对插入片段测序,进行遗传进化分析。结果:确认此次流行株与韩国Seoul/0559/株和美国Shelby株核酸序列相似性最高。结论:国内首次发现诺如病毒GII/12亚型引起急性胃肠炎暴发。     

Detection of a nosocomial outbreak of salmonellosis may be delayed by application of a protocol for rejection of stool cultures

In October 2001 an outbreak of Salmonella enterica serovar enteritidis phage-type 6 occurred in a hospital and a nursing home, both served by the same hospital kitchen. Five nursing home residents died during the outbreak. S. enteritidis was isolated from three of them. Of 231 stool samples from nursing home residents, hospital patients and employees, 82 were culture-positive. All symptomatic patients were treated with oral ciprofloxacin. Inspection of the kitchen showed that during preparation of the desserts implicated in causing the outbreak, temperatures were not measured and storage temperatures were too high. No left-over food samples were available for analysis. According to the 'four-day rule' in use in this hospital, the stool samples related to the first outbreak were not cultured for Salmonella spp., whereas culturing afterwards from both stored specimens and repeats, showed that some of these samples would have been positive for S. enteritidis. Thus without the application of stool culture rejection criteria the outbreak would have been detected one day earlier. With the four-day rule in effect, the outbreak might have been detected much later, if an unusually high number of nursing home residents with gastroenteritis had not been noticed by nursing home physicians. The rule was revised to prevent a possible delay in the future. As a result of this outbreak, the government has announced legislation forbidding the sale of Salmonella-contaminated eggs. An official ban on the use of raw eggs will be included in several hygiene codes.     

Waterborne outbreak of Norwalk-like virus gastroenteritis at a tourist resort,Italy

In July 2000, an outbreak of gastroenteritis occurred at a tourist resort in the Gulf of Taranto in southern Italy. Illness in 344 people, 69 of whom were staff members, met the case definition. Norwalk-like virus (NLV) was found in 22 of 28 stool specimens tested. The source of illness was likely contaminated drinking water, as environmental inspection identified a breakdown in the resort water system and tap water samples were contaminated with fecal bacteria. Attack rates were increased (51.4%) in staff members involved in water sports. Relative risks were significant only for exposure to beach showers and consuming drinks with ice. Although Italy has no surveillance system for nonbacterial gastroenteritis, no outbreak caused by NLV has been described previously in the country.