胶体金法与RT-PCR法测定A群轮状病毒及G血清型分型

目的了解婴幼儿A群轮状病毒腹泻的流行病学特征。方法收集浙江省苍南县人民医院2009年1-12月份的婴幼儿腹泻粪便标本,采用胶体金免疫层析法和逆转录-聚合酶链反应法进行轮状病毒抗原检测和血清型分型,分析G血清型分型。结果 828份婴幼儿腹泻粪便轮状病毒阳性率为33.45%。在轮状病毒腹泻患儿中,1岁以内占49.82%,2岁以内占89.89%。本地区轮状病毒腹泻呈现季节高峰,11月至次年1月为轮状病毒腹泻流行期。毒株分型显示G3型为流行优势株（51.6%）,其次是G1型（26.6%）;另有13.7%为混合感染,包括G1＋G3型（10.5%）、G2＋G3型（2.4%）和G1＋G9型（0.8%）。结论 2009年浙江省苍南县婴幼儿轮状病毒腹泻的主要血清型是G3和G1,G3为优势毒株。 更多还原     

2018-2020年乌鲁木齐市婴幼儿病毒性腹泻监测及病原学分析

分析乌鲁木齐市2018-2020年 < 5岁婴幼儿病毒性腹泻流行病学与病原学特征, 为本地区病毒性腹泻防控提供参考依据。

收集2018-2020年乌鲁木齐市哨点医院门诊 < 5岁婴幼儿病毒性腹泻流行病学信息和粪便标本, 用实时荧光定量PCR方法检测A组轮状病毒(RV)、札如病毒(SAV)、诺如病毒(NV)、肠道腺病毒(EADV)和星状病毒(ASV)核酸, 并用反转录聚合酶链反应(RT-PCR)检测A组RV基因分型。

445份标本中病毒阳性率分别为A组RV 45.02%、NV 10.33%、EADV 9.04%、SAV 5.35%、ASV 1.29%, 混合感染率为28.97%, 7~12月龄组阳性率最高为69.8%, 25~36月龄组最低为2.4%。不同季节A组RV、SAV、ASV阳性率差异有统计学意义(χ²=37.582、21.047、17.685, 均P < 0.001), A组RV 12月份和1月份检出高峰。A组RV的G血清型以G9为主占61.8%, P血清型以P[8]为主占87.7%;NV基因型以GⅡ型为主, 3年检出率分别为93.54%、88.64%、100.00%;病毒性腹泻冬春季高发分别为26.5%、28.5%。

乌鲁木齐市2018-2020年 < 5岁婴幼儿病毒性腹泻中, 2岁以下阳性率最高, A组RV和NV病毒为主要病原。

     

我国1998～1999年流行的婴幼儿腹泻轮状病毒的分型研究

轮状病毒是世界范围内引起婴幼儿腹泻的主要病原。根据病毒外壳蛋白VP4和VP7抗原性的不同可区分为不同型：P（VP4,protease sensitive）型和G（VP7,glycoprotein）型。1998－1999年在中国8个城市（长春、秦皇岛、北京、杭州、福州、广州、成都、昆明）采集了急性腹泻患儿的1093份非细菌性腹泻粪便标本,先进行A组轮状病毒（HRV）的筛选,其中阳性标本433份（39.6％）,电泳型长型占优势（96％）。对HRV标本,再利用血清型特异的MAbELISA和／或RT－PCR进行G分型。结果表明,在1998－1999年,在上述8城市非细菌性腹泻行季节,以HRV G1型为主要流行株,占阳性的83.4％,其次为G3（12.0%）、G4（3.5%)和G2(3.2%）。此外,有3份（0.7％）HRV标本未能分型,12（2.8％）份标本为混合感染,还结合1982－1996年全国12个地区1382份HRV标本的分型资料,分析了我国HVR G血清型的流行规律。实验中又抽样选取了124份GHRV标本,用RT－PCR进行P分型,其中P[8]型76份（61.3％）,P[4]型14份（11.3%）,P[6]型12份(9.7%),P[9]型8份(6.4%)。另外15份（12.1%)未能分出P型,有待进一步检定,实验中HRV分离株除了觉见的P[8]G1(51.4%)、P[4]G2(4.6%)毒株外,还检测到P[8]G3(11.0%)、P[8]G4(6.4%) 和其它较少见的病毒型。以上结果为我国轮状病毒疫苗的应用和开发提供了较系统、清晰的流行病学背景资料。     

昆明婴幼儿腹泻的轮状病毒分子基因特征研究

目的了解昆明婴幼儿腹泻中轮状病毒感染情况。方法收集昆明医学院第一附属医院儿科2004年10-12月住院的腹泻的患儿粪便标本60份,采用聚丙烯酰胺凝胶电泳（PAGE）法和RT—PCR法进行轮状病毒分型。结果60份粪便标本中,29份检测到A组轮状病毒RNA基因,阳性检出率为48.3％,未发现B组及C组轮状病毒。其中RNA长型有29份（100％）,未发现短型和混合型。对29份有明确编号的轮状病毒阳性标本来源的患儿分析显示,平均发病年龄10.5个月。29份标本中,有20份可用RT—PCR分型,均为G3型,未发现其他型。结论A组轮状病毒是昆明地区5岁以下婴幼儿腹泻病的主要病原,3月龄-2岁婴幼儿是轮状病毒的易感人群,以冬季10月份至12月份为流行高峰,基因组以长型为主,血清型为G3型。     

中国河北省卢龙县儿童轮状病毒腹泻研究

轮状病毒是我国儿童重症腹泻的主要病原。按照WHO轮状病毒监测方案,于1999年7月至2003年6月,在河北省卢龙县开展了医院和社区为基础<5岁儿童轮状病毒腹泻的监测。结果表明:卢龙县<5岁儿童腹泻的发病率为1 3次/人/年。4年中全县共有2350名<5岁急性腹泻患儿住院,占所有住院儿童的38%(2350/6213)。住院的腹泻患儿每年有两个高峰,一个是夏季(6～8月),占全年腹泻住院病例的22%;另一个在冬季(12月至次年2月),占58%。住院的轮状病毒腹泻只有一个高峰,是在冬季(12月至次年2月),高峰期的轮状病毒腹泻住院患儿数占全年轮状病毒腹泻患儿住院数的86%。按全年统计,轮状病毒腹泻占住院腹泻患儿的46%,轮状病毒腹泻的住院率为11/1000儿童/年。在门诊腹泻患儿中轮状病毒腹泻患儿占28%,在社区腹泻患儿中轮状病毒腹泻占10%。轮状病毒毒株的分布,G3型(45%)最常见,其次为G1型(35%)、G2型(8%)、G4型(3%)、G9型(0 6%),混合感染较少(1%)。还有8%的毒株未能分出型别。在轮状病毒腹泻患者中,9～11月龄的儿童检出率最高(53%),其次是12～17月龄(51%)、18～23月龄(36%)和6～8月龄(30%)。在4年研究期间共有5～10名1～59月龄儿童可能因为轮状病毒腹泻死亡,其中有1例确诊为P[8]G1型毒株感染。初步估计,该县1～59月龄儿童轮状病毒腹泻的死     

上海部分地区小于5岁儿童轮状病毒及星状病毒性腹泻的临床分析

目的 了解上海地区≤5岁的患急性稀水样便腹泻患儿中轮状病毒感染及星状病毒感染的流行情况及其临床特征。方法 留取2006年6月~2007年3月复旦大学附属儿科医院门诊、住院及医院内感染腹泻患儿的部分粪便标本,应用免疫层析胶体金法检测轮状病毒。排除轮状病毒感染后,应用酶联免疫吸附试验(ELISA)检测星状病毒。本实验主要研究对象为年龄≤5岁,病程≤2周,大便培养无条件致病菌生长患儿。结果 共收集724例急性腹泻粪便标本,年龄≤5岁人群轮状病毒阳性检出率42.5%,约85%患儿年龄≤2岁;病例全年均有发生,发病高峰主要集中在2006年12月~2007年1月。共240例急性腹泻轮状病毒阴性粪便标本中,年龄≤5岁人群星状病毒阳性检出率11.6%,53.6%患儿年龄≤2岁;观测期间病例散发,发病高峰主要集中在2006年10月~2007年1月。结论 轮状病毒是上海地区婴幼儿病毒性腹泻的重要病原,部分患儿伴肠道外损伤。星状病毒是上海地区婴幼儿病毒性腹泻的又一重要病原。     

南京地区2012～2013年G9型轮状病毒VP7基因特征的分析

A组轮状病毒是引起成人和婴幼儿急性腹泻的主要病原。了解轮状病毒流行株的型别,对主要中和抗原VP7的编码基因进行遗传变异分析,可为当地轮状病毒疫苗的应用和开发提供指导。我们对2012年10月至2013年12月南京地区908例腹泻门诊患者的粪便标本进行轮状病毒检测,采用RT-PCR方法对随机抽取的50份阳性标本进行G分型,并对其中19份G9型轮状病毒的VP7基因序列测序分析。结果发现轮状病毒阳性率11.34%(103/908),其中以G9型为主,占78.0%(39/50),其次是G2、G1和G3型。对G9型轮状病毒VP7基因核苷酸序列进行分析,显示主要分为G9-VI亚型和G9-III亚型,以G9-VI亚型为主,且属于中国和日本G9型轮状病毒亚簇,部分毒株在A、B、C、F四个中和抗原表位中有变异,这可能有助于G9型轮状病毒的流行,值得引起注意。     

婴幼儿腹泻A群轮状病毒G和P的基因分型研究

目的研究浙江萧山医院婴幼儿童腹泻标本中人轮状病毒（Human Rotavims）毒株的感染情况及G和P基因型流行特点。方法收集该院2009年8月至2010年8月腹泻儿童15233份粪便标本采用酶联免疫吸附试验、逆转录-巢式聚合酶链反应进行轮状病毒病原检测,将128份阳性标本进行VP7和VP4基本分型。结果15233份婴幼儿腹泻标本中有2706份标本为轮状病毒阳性,阳性率17．8％;男孩和女孩检出率差异无统计学意义,以6-12月龄段检出率最高;对128份阳性标本进行G血清分型和P基因分型,G1型53份（41．4％）、G3型38份（29．7％）、G1G3型17份（13．3％）、G未分型20份（15．6％）;P[8]型72份（56．3％）、P[4]型16份（12．5％）、P[8]P[4]型3份（2．3％）、P未分型37份（28．9％）,G血清型和P基因型的组合以G1P[8]为主,占29．7％（38／128）。结论浙江萧山医院A群轮状病毒G血清以G1型为主,其次为G3型,P基因型以P[8]型为主。     

重庆地区成人感染性腹泻病原学分析

目的 研究重庆地区成人感染性腹泻患者的病原学特点.方法 采用MICROSCAN系统进行粪便细菌培养,用ELISA和多重PCR方法进行病毒检测.结果 130例腹泻标本中,检出细菌阳性17例,包括沙门菌属7例,志贺菌属5例,副溶血弧菌3例和嗜水气单胞菌2例.病毒检测中,单重感染30例,双重感染10例,三重感染1例.A组轮状病毒检出率为3.85％ (5/130),B轮状病毒检出率为3.85％ (5/130),C组轮状病毒检出率为15.4％ (20/130),诺如病毒检出率为9.23％ (12/130),星状病毒检出率为4.62％(6/130),札如病毒检出率为3.08％ (4/130),腺病毒检出率为0.77％(1/130).结论 重庆地区成人腹泻患者中,沙门菌和志贺菌为细菌感染的主要菌种,轮状病毒和诺如病毒为病毒感染的主要病原体.     

杭州地区腹泻儿童轮状病毒与腺病毒感染调查

目的 分析不同季节及不同年龄段腹泻患儿感染轮状病毒（RV）与腺病毒（Eads）情况,为临床提供早期、快速、准确、可靠的依据,以便及时采取相应的治疗使患儿早日康复。方法 采用杭州艾博医药有限公司提供的轮状病毒（A组）/腺病毒检测试剂盒（乳胶法）,对2012年1月至2015年12月浙江省人民医院门诊和住院就诊的3 368例腹泻儿童粪便标本进行轮状病毒和腺病毒抗原检测。结果 3 368例腹泻儿童粪便中,RV阳性578例占17.16%,Eads阳性106例占3.15%。11月至次年1月为轮状病毒感染的高发月份,1～3岁为儿童感染的高发年龄;腺病毒感染阳性率低,呈散发性。结论 杭州地区近年1～3岁婴幼儿的腹泻主要是由轮状病毒和腺病毒引起,其中轮状病毒感染率明显高于腺病毒,且有明显的季节性;而腺病毒感染呈散发流行,未表现出明显的季节性及年龄分布。     

Acute gastroenteritis and enteric viruses in hospitalised children in southern Brazil: aetiology,seasonality and clinical outcomes

Viral acute gastroenteritis (AG) is a significant cause of hospitalisation in children younger than five years. Group A rotavirus (RVA) is responsible for 30% of these cases. Following the introduction of RVA immunisation in Brazil in 2006, a decreased circulation of this virus has been observed. However, AG remains an important cause of hospitalisation of paediatric patients and only limited data are available regarding the role of other enteric viruses in these cases. We conducted a prospective study of paediatric patients hospitalised for AG. Stool samples were collected to investigate human adenovirus (HAdV), RVA, norovirus (NoV) and astrovirus (AstV). NoV typing was performed by nucleotide sequencing and phylogenetic analysis. From the 225 samples tested, 60 (26%) were positive for at least one viral agent. HAdV, NoV, RVA and AstV were detected in 16%, 8%, 6% and 0% of the samples, respectively. Mixed infections were found in nine patients: HAdV/RVA (5), HAdV/NoV (3) and HAdV/NoV/RVA (1). The frequency of fever and lymphocytosis was significantly higher in virus-infected patients. Phylogenetic analysis of NoV indicated that all of these viruses belonged to genotype GII.4. The significant frequency of these pathogens in patients with AG highlights the need to routinely implement laboratory investigations.     

Continuous detection of norovirus and astrovirus in wastewater in a coastal city of China in 2014–2016

Norovirus (NoV) and human astrovirus (HAstV) are important causative agents of acute gastroenteritis in children and adults. They are comprised of multiple genotypes and attention should be paid to genotype changes or emergence of new genetic variants. To study the prevalence and diversity of NoV GI, GII, and HAstV circulating in eastern China, we conducted a three-year environmental surveillance in a coastal city of Yantai. Thirty-six sewage samples were collected, processed, and examined for the presence of viral genomes by PCR. The results showed that NoV GI, GII, and HAstV were detected in all 36 samples. Six NoV GI genotypes, 11 NoV GII genotypes, and 5 HAstV serotypes were identified; GI.6, GII.17, and HAstV-5 were the most prevalent types, respectively. Persistent existence of NoV GII.17 Kawasaki 308 variant was observed during whole study period. Phylogenetic analysis reflected multiple transmission lineages in local population for both viruses. Our results reflect continuous presence of enteric viruses in sewage, improve our understanding on their molecular epidemiology, and demonstrate surveillance on sewage is an effective approach in understanding the local circulation of enteric viruses.     

Low prevalence of rotavirus and high prevalence of norovirus in hospital and community wastewater after introduction of rotavirus vaccine in Nicaragua

Rotavirus (RV) and norovirus (NoV) are major causes of pediatric diarrhea and are altogether associated with approximately 800,000 deaths in young children every year. In Nicaragua, national RV vaccination program using the pentavalent RV5 vaccine from Merck was implemented in October 2006. To determine whether RV vaccination decreased the overall number of RV infections, we investigated the occurrence of RV and NoV in wastewater in the city of León from July 2007 to July 2008 and compared these data with pre-vaccination data. The major finding was the low prevalence of RV compared to NoV in all sampling points (11% vs 44%, p<0.05), and that RV concentration was lower as compared to NoV. RV was observed mainly during the rainy season (July-September), and the majority of all RV detected (6/9) belonged to subgroup (SG) I. The partial VP7-gene obtained from one RV positive sample was similar (99% nt identity) to a G6 VP7-gene of bovine origin and similar to the corresponding gene of the vaccine strain (98%). Furthermore RV G-types 2 and 4 were found in the incoming wastewater. NoV strains were detected throughout the year, of which a majority (20/21) were of genotype GII.4. We conclude that the introduction of RV vaccination reduced the transmission of RV in the community in Nicaragua. However, the burden of diarrhea in the country remains high, and the high prevalence of NoVs in hospital and municipal wastewater is noteworthy. This study highlights the need for further assessment of NoV following RV vaccine introduction.     

小儿病毒性腹泻的临床预后及危险因素分析

目的:探讨小儿病毒性腹泻的临床预后及危险因素。方法:纳入病例是2018年1月至2019年12月本院收治的200例小儿病毒性腹泻患者,回顾性分析其临床资料,分析小儿病毒性腹泻的临床特点。根据疾病痊愈情况分组,将128例痊愈患者作为实验组,72例未痊愈患者作为参照组,Logistic分析小儿病毒性腹泻患者疾病痊愈的危险因素。结果:200例患者中,HAstV感染121例,占60.50%;HAD感染36例,占18.00%;HUCV感染15例,占7.50%;HRV感染10例,占5.00%;混合感染18例,占9.00%。HAstV类型病毒性腹泻患者临床症状以呼吸道症状、发热、呕吐为主;HAD类型病毒性腹泻患者临床症状以发热、呕吐为主;HUCV类型病毒性腹泻患者临床症状以呼吸道症状、发热为主;HRV类型病毒性腹泻患者临床症状以发热、呕吐为主;混合感染类型病毒性腹泻患者临床症状以呼吸道症状、发热、呕吐为主。年龄、中枢神经损害、心肌损害、近1 w是否接触过腹泻是小儿病毒性腹泻患者疾病痊愈的危险因素,P0.05。结论:小儿病毒性腹泻不同感染类型,临床症状各不相同。小儿病毒性腹泻患者预后与年龄、中枢神经损害、心肌损害、近1 w是否接触过腹泻有着极为密切的联系,应当引起临床重视与关注。     

Gastrointestinal Infections and Diarrheal Disease in Ghanaian Infants and Children: An Outpatient Case-Control Study

IntroductionDiarrheal diseases are among the most frequent causes of morbidity and mortality in children worldwide, especially in resource-poor areas. This case-control study assessed the associations between gastrointestinal infections and diarrhea in children from rural Ghana.MethodsStool samples were collected from 548 children with diarrhea and from 686 without gastrointestinal symptoms visiting a hospital from 2007–2008. Samples were analyzed by microscopy and molecular methods.ResultsThe organisms most frequently detected in symptomatic cases were Giardia lamblia, Shigella spp./ enteroinvasive Escherichia coli (EIEC), and Campylobacter jejuni. Infections with rotavirus (adjusted odds ratio [aOR] = 8.4; 95% confidence interval [CI]: 4.3–16.6), C. parvum/hominis (aOR = 2.7; 95% CI: 1.4–5.2) and norovirus (aOR = 2.0; 95%CI: 1.3–3.0) showed the strongest association with diarrhea. The highest attributable fractions (AF) for diarrhea were estimated for rotavirus (AF = 14.3%; 95% CI: 10.9–17.5%), Shigella spp./EIEC (AF = 10.5%; 95% CI: 3.5–17.1%), and norovirus (AF = 8.2%; 95% CI 3.2–12.9%). Co-infections occurred frequently and most infections presented themselves independently of other infections. However, infections with E. dispar, C. jejuni, and norovirus were observed more often in the presence of G. lamblia.ConclusionsDiarrheal diseases in children from a rural area in sub-Saharan Africa are mainly due to infections with rotavirus, Shigella spp./EIEC, and norovirus. These associations are strongly age-dependent, which should be considered when diagnosing causes of diarrhea. The presented results are informative for both clinicians treating gastrointestinal infections as well as public health experts designing control programs against diarrheal diseases.     

Prevalence and Genetic Diversity of Enteric Viruses in Children with Diarrhea in Ouagadougou,Burkina Faso

Enteric viruses are a major cause of diarrhea in children, especially those under five years old. Identifying the viral agents is critical to the development of effective preventive measures. This study aimed to determine the prevalence and genetic diversity of common enteric viruses in children under five years old in Burkina Faso. Stool samples from children with (n = 263) and without (n = 50) diarrhea disorders were collected in Ouagadougou, Burkina Faso from November 2011 to September 2012. Rotavirus, norovirus, sapovirus, astrovirus, adenovirus and Aichivirus A were detected using real-time or end-point (RT-)PCR. Rotavirus strains were G and P genotyped by multiplex RT-PCR and other viral strains were characterized by sequencing of viral subgenomic segements. At least one viral agent was detected in 85.6% and 72% of the symptomatic and asymptomatic patients, respectively. Rotavirus (63.5%), adenovirus (31.2%) and genogroup II norovirus (18.2%) were the most prevalent viruses in symptomatic patients, but only rotavirus and genogroup II norovirus were significantly associated with diarrhea (OR: 7.9, 95%CI: 3.7–17; OR: 3.5, 95%CI: 1–11.7, respectively). Sapovirus (10.3%), astrovirus (4.9%), genogroup I norovirus (2.7%) and Aichivirus A (0.8%) were less prevalent. The predominant genotype of rotavirus was G9P[8] (36.5%), and the predominant norovirus strain was GII.4 variant 2012 (71.4%). Among sapovirus, the genogroup II (87.5%) predominated. Astrovirus type 1 (41.7%) was the most frequent astrovirus identified. Aichivirus A belonged to the three genotypes (A, B and C). Enteric adenoviruses type 40 and 41 were identified in 10.2% and 5.1% respectively. Several cases of co-infections were detected. The results highlight the high prevalence and the high diversity of enteric viruses in Burkinabe children.