浙江省2008-2009年三起诺如病毒胃肠炎暴发的病原分子特征分析

Objective To study the molecular characteristic of norovirus in 3 outbreaks of gastroenteritis in Zhejiang province. Methods During January 2008 and December 2009, fecal specimens of patients were collected from 3 outbreaks of acute viral gastroenteritis. Noroviruses were detected by Real-time RT-PCR. Part of the positive samples were randomly selected and detected by RT-PCR. PCR products were sequenced. Sequence analysis was undertaken based on partial sequence of RNA dependent RNA polymerase(RdRp)and capsid protein gene. Some positive samples were amplified by 3' RACE(rapid amplification of cDNA 3' ends), 3200 bp in length. The exact whole ORF2, ORF3 and 3' untranslation regions(UTR)gene of norovims were identified. Results There were in total 3 outbreaks of viral gastroenteritis caused by norovirus being reported. A total of 62 stools were obtained from cases with acute gastroentefitis. Noroviruses were detected in 41 cases including 27 strains of genogroup Ⅰ norovirus and 9 strains of genogroup Ⅱ norovirus, 5 strains of genogroup Ⅰ + Ⅱ norovirus. Four genotypes including G Ⅰ .8, G Ⅱ .b, G Ⅰ .2/0 Ⅰ .6 recombination together with co-infection of G Ⅰ .8 and G Ⅱ .b were detected. Conclusion Norovirus was confirmed as the major cause of outbreaks of viral gastroenteritis in Zhejiang province and multiple genotype of norovirus were identified from the outbreaks. It was the first time to have found a recombinant of G Ⅰ .6 capsid and G Ⅰ .2 polymerase norovims as well as the co-infection of G Ⅰ .8 and G Ⅱ .b norovirus in the same sample.     

浙江省2008-2009年三起诺如病毒胃肠炎暴发的病原分子特征分析

Objective To study the molecular characteristic of norovirus in 3 outbreaks of gastroenteritis in Zhejiang province. Methods During January 2008 and December 2009, fecal specimens of patients were collected from 3 outbreaks of acute viral gastroenteritis. Noroviruses were detected by Real-time RT-PCR. Part of the positive samples were randomly selected and detected by RT-PCR. PCR products were sequenced. Sequence analysis was undertaken based on partial sequence of RNA dependent RNA polymerase(RdRp)and capsid protein gene. Some positive samples were amplified by 3' RACE(rapid amplification of cDNA 3' ends), 3200 bp in length. The exact whole ORF2, ORF3 and 3' untranslation regions(UTR)gene of norovims were identified. Results There were in total 3 outbreaks of viral gastroenteritis caused by norovirus being reported. A total of 62 stools were obtained from cases with acute gastroentefitis. Noroviruses were detected in 41 cases including 27 strains of genogroup Ⅰ norovirus and 9 strains of genogroup Ⅱ norovirus, 5 strains of genogroup Ⅰ + Ⅱ norovirus. Four genotypes including G Ⅰ .8, G Ⅱ .b, G Ⅰ .2/0 Ⅰ .6 recombination together with co-infection of G Ⅰ .8 and G Ⅱ .b were detected. Conclusion Norovirus was confirmed as the major cause of outbreaks of viral gastroenteritis in Zhejiang province and multiple genotype of norovirus were identified from the outbreaks. It was the first time to have found a recombinant of G Ⅰ .6 capsid and G Ⅰ .2 polymerase norovims as well as the co-infection of G Ⅰ .8 and G Ⅱ .b norovirus in the same sample.     

诺如病毒感染与致病机制研究进展

<正>诺如病毒(norovirus,NV)是引起人非细菌性急性胃肠炎疫情暴发和婴幼儿急性腹泻的主要病原体,特别是引起病毒性腹泻暴发的重要病因。诺如病毒可分为7个基因组(GⅠ～GⅦ组),40个基因型。GⅠ、GⅡ、GⅣ组主要引起人类感染,其中GⅠ组有9个基因型,GⅡ有22个基因型,而GⅣ仅有2个基因型。从20世纪90年代至今,GⅡ.4 NV已经逐渐成为了世界范围内的优势流行株,并且造成了至少4次全球性的疾病流行。长期以来,普遍存在着诺如病毒优     

浙江省2006-2007年暴发性胃肠炎中诺如病毒的分子流行病学分析

目的 分析浙江省2006-2007年暴发性病毒性胃肠炎中诺如病毒感染的分子流行病学特征.方法 收集监测期间暴发性病毒性胃肠炎疫情患者的粪便标本及相关流行病学资料.采用RT-PCR及荧光定量RT-PCR检测诺如病毒.随机选掸部分阳性标本扩增部分多聚酶区和衣壳蛋白片段,进行序列测定,结合诺如病毒I(GI)、Ⅱ(GⅡ)基因型参考株进行进化分析.结果 诺如病毒感染暴发性胃肠炎共5起,发生时间均集中于9-12月,送检标本63份,诺如病毒检测阳性45份.序列分析结果显示,浙江省诺如病毒序列与诺如病毒GⅡ/4型参考株同源性最高,其中与北京2006年、荷兰2006年诺如病毒GⅡ/4型变异株最为接近,核苷酸同源性分别为99.7%和98.5%～99.0%.诺如病毒与北京、荷兰流行的GⅡ/4型变异株处于同一分支.结论 诺如病毒是浙江省病毒性腹泻暴发疫情的重要病原体,流行时间集中于秋季,其流行株为GⅡ/4型变异病毒株.     

云南省某中学一起诺如病毒暴发疫情的病原鉴定和基因特征分析

目的明确2022年云南省某中学一起急性肠胃炎暴发疫情的病原和基因特征。方法采集这起暴发疫情中的学生和厨工病例的肛拭子, 使用诺如病毒GⅠ/GⅡ双通道核酸检测试剂盒定性检测。GⅠ和GⅡ阳性的样本分别采用引物G1SKF/G1SKR和MON431/G2SKR进行病毒扩增和测序, 通过在线分型工具及系统进化分析确定病毒基因型别。结果共采集58份肛拭子, 实验室检测阳性率为53.45%(31/58)。其中, GⅠ阳性样本22份, GⅡ阳性样本14份, GⅠ和GⅡ合并感染样本5份。31份阳性样本基因测序获得毒株序列22株, 包括诺如病毒GⅠ型15株, 其中GⅠ.6[P6]型9株、GⅠ6.[P11]型6株;诺如病毒GⅡ.4[P16]型7株。结论本起疫情由诺如病毒GⅠ.6[P6]型、GⅠ.6[P11]型和GⅡ.4[P16]型混合感染引起, 这是云南省首次在诺如病毒暴发疫情中检出GⅠ型。     

深圳市罗湖区2018年1月——2021年10月诺如病毒流行病学分析

目的 分析深圳市罗湖区2018年1月—2021年10月诺如病毒的病原学型别及流行状况,初步预测流行趋势,为诺如病毒防控提供有效依据。方法 采用real-time PCR技术对采集的标本进行病毒鉴定,利用基因测序对诺如病毒阳性样本进行序列分析。结果 2018年1月—2021年10月共监测散发病毒性腹泻440例,其中诺如病毒引发的占17.27%,同期聚集性腹泻疫情一共54宗,诺如病毒引发占比85.19%,主要流行的基因亚型为GⅡ．2、GⅡ．4、GⅡ．17。结论 诺如病毒是病毒性腹泻的主要病原体,聚集性腹泻基本由其引发。辖区流行的诺如病毒仍以GⅡ为主,但基因亚型中GⅡ．2占比增多,需加大对其监测,防止大规模疫情暴发。     

一起两校同源诺如病毒急性胃肠炎暴发调查

诺如病毒(norovirus,NV)为人类杯状病毒(human caliciviruses,HuCV),是引起病毒性腹泻最常见的病原体之一[1-2].浙江省近年来已有多起诺如病毒引起暴发疫情的报道[1,3-5,给全省带来了较大疾病负担.2012年11月23日,浙江省临安市浙江农林大学报告一起学生恶心、呕吐、腹痛、腹泻事件,次日,湖州市德清县高级中学也出现症状类似的暴发疫情,经实验室初步检测核实,均为诺如病毒暴发疫情.经现场调查,结果报道如下.     

佛山市南海区诺如病毒引起急性胃肠炎暴发的流行病学研究

目的 了解佛山市南海区暴发的急性胃肠炎中诺如病毒感染的流行特征.方法 对南海区 2006-2007 年暴发的2起急性病毒性胃肠炎疫情进行流行病学调查和分子流行病学研究,将收集到的患者的粪便、肛拭子标本,采用针对诺如病毒的特异性引物和逆转录聚合酶链反应(RT-PCR)技术进行检测,再经过分析核酸序列分析病毒的基因型.结果 两起暴发流行的急性胃肠炎均由GⅡ.4亚型的诺如病毒引起.结论 诺如病毒是引起南海区秋冬季急性胃肠炎暴发流行的主要病原体,且引起暴发的流行病毒株属于GⅡ.4型.     

GGⅠ诺如病毒致胃肠炎饮水型暴发的调查

诺如病毒是引起人类急性胃肠炎的重要病原体,其感染基因型包括GG Ⅰ、GG Ⅱ和GGⅣ[1].调查显示我国人群诺如病毒感染十分普遍[2],尽管如此,国内报道的诺如病毒基因型多为GGⅡ[3-5],而GGⅠ特别是多区域人群暴发的研究较少涉及.2008年3月杭州市某区发生一起GG Ⅰ诺如病毒的暴发疫情,调查结果如下.     

广东省2008－2009 年沙门菌监测

目的 了解广东省腹泻患者中沙门菌的感染及沙门菌暴发的情况以及沙门菌株的血清型别、耐药性和分子特征.方法 对纳入研究的腹泻病患者进行沙门菌的检测,对日常监测中分离到的菌株和暴发监测收集到的菌株进行血清分型、药物敏感试验和脉冲场凝胶电泳(PFGE)分型.结果 2008年共检测1922份粪便标本,分离到7l株沙门菌,阳性率为3.7%;2009年检测2110份粪便标本,分离到85株沙门菌,阳性检出率为4.0%;156株菌共分37种血清型,鼠伤寒和肠炎沙门菌居多;监测到10起由沙门菌污染引起的食物中毒事件,其中有4起由肠炎沙门菌引起,有3起由鼠伤寒沙门菌引起;发现1起疑似肠炎沙门菌暴发,并开展流行病学调查,结果提示4名病例中有2名病例是感染同一来源的肠炎沙门菌;229株沙门菌对头孢类和喹诺酮类抗菌药物敏感率达80%以上,59.3%是多重耐药沙门菌.结论 在广东省引起感染性腹泻和食物中毒的沙门菌主要为肠炎沙门菌和鼠伤寒沙门菌.

Abstract：

Objective To understand the infection of Salmonella (S.) in patients with diarrhea and outbreaks caused by Salmonella to identify the serotypes, resistance to antibiotics and PFGE types of the strains from the surveillance program in Guangdong province. Methods S. strains from patients with diarrhea were detected, and all the positive strains collected in routine and outbreak surveillance programs, were tested by serum agglutination, antibiotic susceptibility and PFGE.Results 71 S. strains were isolated from 1922 stool samples in 2008, with positive rate as 3.7%.85 S. strains were isolated from 2110 stool samples in 2009, with positive rate as 4.0%. All the 156 strains were divided into 37 serotypes, with S. serotype typhimurium and enteritidis as the most common serotypes. 10 incidents of food poisoning were detected, of which 4 were caused by enteritidis and 3 by typhimurium. A suspected outbreak by enteritidis was discovered and under epidemiological investigation. The findings indicated that 2 of the 4 patients from this outbreak were infected with identical enteritidis isolates. 80% of the 229 isolates were found susceptible to cephalosporins and quinoione and 59.3% of them were muitiresistant to the antibiotics. Conclusion S. enteritidis and S. typhimurium were the most common serotypes that caused infectious diarrhoea and food poisoning in Guangdong province.     

中国2006—2007年诺如病毒胃肠炎暴发及其病原学特征分析

目的 了解中国2006-2007年诺如病毒胃肠炎暴发及其病毒基因型别.方法 收集2006-2007年19起胃肠炎暴发的流行病学资料以及腹泻粪便样本,用RT-PCR方法对201份粪便标本进行诺如病毒、轮状病毒、星状病毒、腺病毒和札如病毒检测,对扩增产物进行序列测定.用Clustal X 1.83和MEGA4.0生物软件对诺如病毒序列进行序列比对和系统进化分析.结果 诺如病毒是引起病毒性胃肠炎暴发的主要病原之一(12/19,63.2%).在12起诺如病毒胃肠炎暴发中,变异株GⅡ-4/2006b是引起暴发的流行优势株(11/12,91.7%),其他型别包括GⅡ-17、GⅡ-6和GⅡ-3.诺如病毒引起的疫情暴发主要集中在冬春季节(12月至4月),发生场所多样,且各年龄组人群均有发病.同时,12起诺如病毒暴发中有2起与其他腹泻病毒混合感染,且在同一起暴发中病原具有病毒多样性和基因型别多样性.结论 诺如病毒是2006-2007年引起胃肠炎暴发疫情的主要病原之一,变异株GⅡ-4/2006b为流行优势株.     

杭州地区2009-2010年急性腹泻患者中杯状病毒的检测和分型

目的 了解2009-2010年杭州地区急性腹泻患者中杯状病毒基因型别和分子流行病学特征.方法 收集2009-2010年杭州地区920例急性腹泻患者的粪便标本和流行病学资料,用多重PCR方法进行杯状病毒检测,测定部分标本的阳性扩增片段基因序列,并对序列进行系统进化分析.结果 急性腹泻患者杯状病毒检出率为21.8％(201/920),其中诺如病毒(NV)G Ⅰ型25例、GⅡ型170例、札如病毒(SAV) 11例,4例为NVⅠ型和Ⅱ型混合感染、1例为NVⅡ型和SAV混合感染.NV基因型别包括:GⅠ-1(3株)、G Ⅰ-2(1株)、GⅡ-4/2006b变异株(7株)、GⅡ-2(1株)、GⅡ-7(1株)和GⅡ-4/2008变异株(2株);SAV基因型别包括:G Ⅰ -2(5株)、GⅠ-1(4株)和GⅡ-1(1株).杯状病毒的流行在不同季节、年龄组人群均有发病.结论 杯状病毒是2009-2010年引起杭州地区急性腹泻的主要病原之一,其病原具有病毒多样性和基因型别多样性,NVGⅡ-4/2006b变异株或类似株可能是2009-2010年杭州地区流行的优势株.     

GⅡ-4型诺如病毒ORF2基因克隆及序列分析

目的 获得中国流行株GⅡ-4型诺如病毒(NoV)ORF2基因序列,为进一步研究诊断、疫苗提供基础依据.方法 从GⅡ-4型NoV粪便中提取病毒RNA,经逆转录后PCR并克隆到simple PMD18-T载体获得完整开放读码柜架(ORF2)阅读框序列.结果 获得了正确序列的基因Ⅱ-4,亚型(genogroup Ⅱ-4,GⅡ-4)NOV ORF2基因序列,经软件分析,3株NOV ORF2序列编码的氨基酸大小约为54KD,与VA387(GⅡ-4)氨基酸同源性为92%～93%.ORF2编码的氨基酸序列存在3个相对保守序列.结论 获得GⅡ-4型NoV ORF2基因序列,为进一步表达病毒衣壳蛋白及Nov感染的诊断试剂及防治疫苗药物研制提供基础依据.     

Outbreaks of noroviral gastroenteritis in Florida, 2006-2007

Noroviruses are an important cause of sporadic cases and outbreaks of acute gastroenteritis. During 2006-2007, widespread increases in acute gastroenteritis outbreaks consistent with norovirus were observed in the United States. We conducted a statewide survey to characterize norovirus outbreak activity in Florida during a 1-year period. From July 2006 to June 2007, 257 outbreaks of norovirus gastroenteritis were identified in 39 of Florida's 67 counties. About 44% of outbreaks were laboratory confirmed as norovirus and 93% of these were due to genogroup GII. About 63% of outbreaks occurred in long-term care facilities and 10% of outbreaks were classified as foodborne. The median number of ill persons per outbreak was 24, with an estimated total of 7880 ill persons. During the study period, norovirus outbreak activity in Florida was widespread, persistent, and consistent with increased activity observed in other parts of the country.     

牡蛎和粪便中GⅠ、GⅡ型诺如病毒实时聚合酶链反应检测方法的建立

目的 建立适用于牡蛎和粪便中快速、特异、灵敏的GⅠ、GⅡ型诺如病毒(NV)定量分型诊断方法.方法 通过对GⅠ、GⅡ型NV基因组保守序列的比对分析,设计高度特异的引物和探针,建立以TaqMan探针为基础的实时聚合酶链反应方法(TaqMan Real-time RT-PCR).结果 该方法对NV核酸检测高度特异,且GⅠ和GⅡ型之间无交叉反应,最低检出限达102 copies/μl.对90份新鲜牡蛎样品和37份腹泻粪便标本分别用常规RT-PCR和笔者建立的TaqMan Real-time RT-PCR进行NV检测,发现牡蛎样品中后者的检出率明显高于前者,而粪便标本中两者无明显差别.同时对阳性标本的测序分析证实结果准确可靠.结论 研究中建立的TaqMan Real-time RT-PCR方法可用于海产品标本及粪便中NV定量及分型检测,可作为应对NV胃肠炎暴发的有效诊断方法.     

南宁市朝阳溪污水中诺如病毒GⅡ.4型变异株基因分析

目的了解南宁市朝阳溪污水中诺如病毒(Norovirus,NV)GⅡ.4变异株的基因特征。方法 2011年1~12月采集72份南宁市朝阳溪污水,经实时荧光逆转录-聚合酶反应(Realtime RT-PCR)方法进行NV核糖核酸(RNA)检测,阳性标本进行基因克隆测序和遗传进化分析。结果 72份污水中均能检测到NV RNA,基因Ⅰ组(GenogroupⅠ,GⅠ)NV RNA阳性率为70.83%,基因Ⅱ组(GenogroupⅡ,GⅡ)阳性率则达100.00%,共为16个基因型,有18株为GⅡ.4型,其中2株为2006b变异株,其余16株均为2010变异株。检测到的2006b变异株和2010变异株在衣壳蛋白N/S区第94位氨基酸均发生变异,而在第6位、第15位仅2010变异株发生了变异,2006b变异株(除SW2011041-2株外)均无变异。这些变异株与同年南宁市腹泻病例中NV GⅡ.4型变异株核苷酸型内同源性分别为96.70%~98.93%和97.42%~99.64%。结论南宁市朝阳溪污水中NV存在很高的遗传多样性,其来源可能是周边居民中的感染者;GⅡ.4型NV中2010变异株为优势株,其在人群中可能存在隐性感染;污水监测是人间NV感染监测的重要补充。     

苏州市14起诺如病毒胃肠炎暴发疫情流行特征分析

目的了解苏州市诺如病毒胃肠炎暴发疫情的流行病学特征。方法对2010-2014年发生的14起诺如病毒胃肠炎暴发疫情进行描述性流行病学分析,采用聚合酶链反应对病例肛拭子、粪便、呕吐物、水、环境涂抹标本进行核酸检测。结果 2011-2014年,苏州市共报告14起诺如病毒胃肠炎暴发疫情,罹患率波动范围为0.89%~21.18%。14起疫情中12起(85.71%)发生于托幼机构和中小学校,医院、农村各1起。全年均有发生,但以寒冷季节发生较多。仅1起查明与二次供水受到污染有关,13起未证实暴发原因。核酸检测结果12起由诺如病毒GII基因组引起,2起由GI基因组引起。结论诺如病毒已成为苏州市急性胃肠炎暴发疫情的重要病原,托幼机构和学校是高发场所,GII基因组是主要基因组。在暴发疫情处理中,应加强传染源和传播途径的调查。     

国内诺如病毒胃肠炎疫情分子流行病学分析

目的 研究诺如病毒聚集性胃肠炎疫情在国内的流行现状. 方法 运用系统综述的方法在万方数据库、维普咨讯数据库、CNKI数据库、生物文献数据库及pubmed数据库中检索关于国内诺如病毒聚集性胃肠炎疫情的文献.结果 国内从2003年1月-2012年6月共发生83起诺如病毒胃肠炎疫情.从地点分布来看,疫情主要集中在广东省、浙江省,其次是北京市、广西省;从季节分布来说,诺如病毒胃肠炎疫情每季度均有发生,但秋冬季高发（10、1、2月份）;从发生场所来说,疫情主要发生在学校、医疗卫生保健机构;水源性、人与人密切接触传播为其主要传播途径.人与人密切接触传播多为GⅡ.4型感染所致. 结论 学校等作为诺如病毒胃肠炎疫情防控工作的重点场所,应加强诺如病毒防治知识培训,提高疫情报告敏感性和及时性.     

Norovirus and gastroenteritis in hospitalized children, Italy

Noroviruses were detected in 48.4% of 192 children (<3 years of age) hospitalized for gastroenteritis in Palermo, Italy, during 2004; predominant genotypes were GGIIb/Hilversum and GGII.4 Hunter. Of children with viral enteritis, 19.6% had a mixed norovirus-rotavirus infection. The severity of infection was lower for norovirus than for rotavirus but increased in co-infection.