诺沃克样病毒广州地区分子流行病学分析

目的根据广州市多起由诺沃克样病毒引起的食物中毒患者粪便标本中检测到的Ⅱ型诺沃克样病毒序列,初步了解广州地区诺沃克样病毒的分子流行病学情况。方法用RT-PCR对病人粪便中的诺沃克样病毒核酸进行扩增,将阳性产物测序,并与GenBank中的序列相比较。结果归纳出4个代表性序列,并且相似率都不高。结论我国存在诺沃克样病毒引起的食物中毒,而其RNA复制酶的保守区段变异普遍较大。     

一起医院内诺如病毒感染性腹泻事件的调查

目的调查分析一起医院内群体性感染性腹泻的原因，为制定防控措施提供科学依据。方法结合现场流行病学调查，采集粪便标本2份、肛拭子标本5份进行肠道致病菌、诺如病毒检测。结果本次疫情流行9d，发病22例（住院病人15例，护工1例，清洁工1例，医务人员5例），罹患率23．91％。2份粪便标本、4份肛拭子标本检出G11型诺如病毒核酸阳性。通过采取隔离治疗感染病例、消毒环境及物品、对医务人员及病人卫生宣教等综合措施，疫情得到及时控制。结论这是一起由诺如病毒感染所致的感染性腹泻事件，密切接触是本起疫情的主要传播途径。     

新型冠状病毒肺炎确诊病例粪便标本的病毒核酸检测

目的了解新型冠状病毒肺炎(COVID-19)确诊病例粪便中2019-nCoV感染情况。方法收集36例确诊病例的粪便标本/肛拭子标本,采用实时荧光RT-PCR检测2019-nCoV载量,利用统计分析软件SPSS 19.0比较病例的带毒率。结果36份标本共检出2019-nCoV核酸阳性20份(55.56%),危重症病例(2/3)和重症病例标本阳性率均为66.67%(6/9)、普通肺炎病例阳性率为62.50%(10/16)、轻症肺炎病例阳性率为25.00%(2/8)。36例确诊病例包括男性22例、女性14例,检出率分别为54.55%和57.14%;病例年龄分布于17~86岁之间,平均为48.75岁。36份标本其中5份肛拭子标本检出阳性标本2份,31份粪便标本检出阳性标本18。临床分型、性别、年龄和标本类型各组间阳性检出率均无统计学差异。结论本研究发现COVID-19患者,无论重症、轻症患者粪便标本均存在2019-nCoV基因检测阳性,提示消化道可能是该病毒排泄渠道,因此可能存在粪-口传播途径,研究结果具有重要的临床和流行病学意义。     

青岛地区腹泻患儿星状病毒感染的分子流行病学特征

目的研究青岛地区婴幼儿腹泻患者星状病毒流行病学特点和基因型别。方法收集青岛地区散发儿童病毒性腹泻患者粪便标本48份,经胶体金法、RT-PCR法分别检测轮状病毒和诺如病毒后,应用RT-PCR进行星状病毒检测,并对阳性标本进行测序,鉴定型别。结果 48份粪便标本中,4份检出星状病毒(8.3%),经序列分析鉴定为HastV-1型。结论青岛地区散发病毒性腹泻患儿存在星状病毒感染,型别为HastV-1。     

90例病毒性胃肠炎患者粪便中诺如病毒检出情况分析

目的 了解病毒性胃肠炎患者中诺如病毒感染情况. 方法 收集北京市2007年2月5~17日共计90例病毒性胃肠炎患者的粪便标本,应用逆转录聚合酶链反应法 (RT-PCR) 和酶联免疫吸附试验 (ELISA) 检测粪便中诺如病毒核酸或抗原,并对RT-PCR阳性标本的PCR产物进行克隆测序. 结果 90例病毒性胃肠炎患者的粪便标本中,35例 (38.89%) RT-PCR为诺如病毒核酸阳性,序列分析结果显示,诺如病毒GⅡ型34例 (97.14%),GⅠ型1例 (2.86%);90例中,39例 (43.33%) 为ELISA检测诺如病毒抗原阳性.以RT-PCR检测结果作为金标准进行比较,ELISA的灵敏度和特异度分别为97.14% (34/35) 和90.91% (50/55) . 结论 诺如病毒是病毒性胃肠炎的主要病原,以GⅡ型流行为主.ELISA快速、简便,其灵敏度和特异度较高,可作为诺如病毒感染的初筛试验.     

一起诺如病毒GⅡ.17型引起的急性胃肠炎病原学诊断及基因特征分析

目的对长沙市2014年12月某厂区暴发的一起急性胃肠炎疫情进行病原学诊断及对其致病原进一步的基因分型研究。方法共采集6例病人肛拭子标本和可疑水源标本4份,提取核酸后,诺如病毒GI/GII型real-time RT-PCR试剂盒检测;阳性标本普通RT-PCR扩增VP1基因片段,产物测序后BLAST比对确定其型别,并构建进化树分析其进化关系。结果 10份标本real-time RT-PCR扩增结果显示为诺如病毒GII型,VP1区片段RT-PCR扩增后产物测序,BLAST比对发现与2014年香港诺如病毒株GII/Hu/HKG/2014/GII.17/CUHK-NS-463同源性最高,达99%,证实为诺如病毒GII.17型;构建系统进化树分析显示本次疫情分离的毒株与日本、香港、台湾等亚洲地区的毒株亲缘关系更为接近,而与美国、法国等地区的毒株亲缘关系则较远。结论诺如病毒是引起此次急性胃肠炎疫情的病原体,且引起暴发的病毒株属于GII.17型。     

RT-PCR和ELISA在检测小儿病毒性腹泻中的应用比较

[目的]RT-PCR和ELISA在检测小儿病毒性腹泻中的效能分析。[方法]收集我院2018年6月～2019年1月在门诊及住院的1个月～5岁400例儿童的腹泻粪便标本。利用RT-PCR和ELISA方法检测标本中的RV、NV、AstV和AdV病毒性病原。具体操作方法,严格按照试剂盒说明书进行操作。[结果]采用RT-PCR检出至少有132份标本含有至少1种病毒核酸:116份单一病毒核酸呈现阳性,14份双重病毒核酸呈现阳性,2份三重以上病毒核酸阳性。核酸阳性的检出率为36.7%。采用ELISA的方法检出98份标本含有至少1种病毒核酸:86份单一病毒核酸阳性,12份双重病毒核酸阳性。核酸阳性的检出率为27.2%。2种检测方法检测RV、NV、AstV和AdV病毒性病原差异有统计学意义。[结论]RT-PCR和ELISA 2种检测方法的检测结果具有很好的一致性。RT-PCR和ELISA此2种检测方法均以RV、NV的阳性检出率较高,RT-PCR法除对EAdV阳性检出率低于ELISA外,对其他3种病毒的检出率均高于ELISA。     

一起由星状病毒1型引起的新生儿腹泻暴发的病原确证

目的 对一起新生儿急性胃肠炎暴发事件进行病原确证.方法 2008年12月至2009年2月,内蒙古自治区某妇幼医院新生儿室发生腹泻流行,高峰期采集38例患儿的45份粪标本,ELISA法检测轮状病毒、腺病毒和星状病毒病原,RT-PCR法检测星状病毒核酸,其中13份星状病毒核酸阳性标本进行测序分析和进化树分析,4份星状病毒及病毒核酸均阳性的标本进行免疫电子显微镜观察.结果 45份粪标本中,轮状病毒、腺病毒病原检测均阴性.30份标本ELISA检测星状病毒病原阳性,阳性率为66.7%;31份标本星状病毒核酸阳性,阳性率为68.9%.采用星状病毒分型引物进行分型,均为星状病毒1型.选择13株与GenBank中星状病毒1型参考株进行比较,其核苷酸序列同源性为90.9%～96.3%.13株星状病毒1型株彼此间核苷酸序列同源性为94.7%～100.0%.随机选择的4份阳性标本,免疫电子显微镜下2份有大量星状病毒颗粒.结论 此起新生儿腹泻暴发由星状病毒1型引起.     

志愿者口服重组诺沃克病毒颗粒(疫苗)的Ⅰ期临床研究

诺沃克病毒隶属杯状病毒科,能引起人类流行性急性胃肠炎。它通过受污染的水或食物以粪-口途径感染人群,流行病学调查显示美国及荷兰90％非细菌性胃肠炎的暴发流行是该病毒引起。由于患者粪便与呕吐物中病毒颗粒含量很低,且缺乏有效的细胞培养系统及动物模型,以及病毒衣壳蛋白很易     

山东半岛地区贝类中诺如病毒污染状况与病毒鉴定初报

目的调查研究山东半岛沿海地区双壳贝类中诺如病毒污状况并对阳性株进行鉴定。方法应用常规RT-PCR与实时荧光定量RT-PCR,对2007年9月到2008年8月间在山东半岛沿海8个地区收集的186份双壳贝类样品进行了诺如病毒的定性和定量检测,并对阳性株核酸片段进行克隆、转化和序列分析,使用DNAStar软件进行核酸序列同源性比较,经MEGA4绘制系统进化树。结果 186份样品中检出5份诺如病毒阳性,检出率为2.67%,诺如病毒含量为大于102拷贝RNA,序列分析与遗传进化树显示5份诺如病毒阳性株均为GGII型NVs,与国内报道的GGII型毒株同源性达到98%以上。结论山东半岛地区贝类中含有的诺如病毒以GGII型为主,冬季检出率较高。     

A food poisoning outbreak caused by purple Washington clam contaminated with norovirus (Norwalk-like virus) and hepatitis A virus

A party of 57 people dined together in a restaurant in Hamamatsu City on December 11, 2001. The next day, 22 of them developed symptoms of acute gastroenteritis, such as diarrhea, vomiting, and fever. Examination of 4 fecal specimens from these patients by ELISA for Norovirus (Norwalk-like virus, NV) detected both genogroup I (GI) and genogroup II (GII) NV in all the 4 specimens. In addition, RT-PCR and real-time PCR methods for NV detected the NV gene. Approximately one month after the outbreak of the food poisoning (acute gastroenteritis) by NV, 4 individuals in the same party developed type A hepatitis. Both RT-PCR and real-time PCR methods for hepatitis A virus (HAV) detected the HAV gene in their fecal specimens. The party of these patients ate purple Washington clam (Saxidomus purpuratus, imported from China) steamed with red pepper. Since this food appeared to have caused the viral infections, the one with the same lot number was subjected to viral examinations, which successfully detected the NV GI, NV GII, and HAV genes. These results led to the conclusion that the clam contaminated with NV and HAV had caused the food poisoning. The DNA sequences of the NV detected in the patients and the clam had 74 to 99% homology, indicating strains of various genotypes. All the strains of HAV that were derived from the patients and the clam were genotype 1A, and these sequences had over 95% homology, but were not completely identical. This outbreak led to the demonstration of imported fishery products as a cause of type A hepatitis, and indicated the need for guiding and enlightening people on the importance of adequate cooking of bivalves.     

Epidemiology of calicivirus infections in Sweden, 1994-1998

Outbreaks of acute gastroenteritis are frequently caused by caliciviruses. Electron microscopy was used to search for these viruses in fecal samples from patients with acute gastroenteritis. Of 5800 samples collected and analyzed from November 1994 to June 1998, 3700 were associated with outbreaks. A total of 676 outbreaks were analyzed, and viruses were found in 67%. Caliciviruses, usually Norwalk-like viruses (NLVs), were found in 407 (89%) of 455 outbreaks, while Sapporo-like viruses were identified in nine outbreaks, including six that were suspected to include foodborne transmission. Sixty percent of the 1041 patients with calicivirus infections were between 70 and 90 years of age. Food- and waterborne infections were associated with 66 calicivirus outbreaks. Virus-positive outbreaks were documented mainly during winter and spring. The longitudinal survey showed that caliciviruses, and especially the NLVs, cause most nosocomial and community-associated outbreaks in Sweden.     

南京地区人感染杯状病毒的病原学检测

目的了解南京地区病毒性腹泻患者中杯状病毒的感染情况,明确该地区杯状病毒的主要基因型和分子进化特点。方法收集南京市儿童医院疑似病毒性腹泻粪便标本155例,南京市某三甲医院成人腹泻标本8例,采用RT-PCR方法检测粪便中诺如病毒和札如病毒的多聚酶区核酸片段,并测序以明确基因型,对核苷酸序列使用CLUSTALW和phylip软件分析病毒的分子进化和亲缘关系。结果婴幼儿的155例标本共检出15例杯状病毒阳性,检出率达到9.7%,成人检出5份阳性,与婴幼儿感染的诺如病毒核酸在多聚酶区未发现显著差异,均为GⅡ4型,与GENBANK上序列号为EU074213(苏州)、EU096514、EF200699的毒株有较高的同源性;婴儿腹泻中有一例经核酸序列比对确认为札如病毒感染,为SGⅡ型,与GENBANK上序列号为AF439862的有94%的同源性。结论杯状病毒是本地区病毒性腹泻的重要病原体,感染的该病毒以诺如病毒GⅡ4型为主,与近年多个地区发现的该病毒有较近的亲缘关系。南京地区存在札如病毒感染。     

Norwalk virus in Norway: an outbreak of gastroenteritis studied by electron microscopy and radioimmunoassay

An outbreak of acute gastroenteritis affected approximately half of 40 children staying at a holiday centre in Southern Norway. By direct electron microscopy Norwalk-like viruses were demonstrated in 4/8 available stool specimens. No other pathogens were detected. Antibody against these viruses was demonstrated by immune electron microscopy in all of 7 convalescent phase sera but in none of 11 acute phase sera collected. Radioimmunoassay examination showed a rise in titre of Norwalk virus antibody in 6 available paired sera. This outbreak of Norwalk virus gastroenteritis in Norway was thus documented by a combined use of direct and immune electron microscopy and radioimmunoassay.