马鞍山、苏州地区5岁以下腹泻儿童轮状病毒G、P分型研究

目的 研究马鞍山和苏州地区5岁以下腹泻儿童轮状病毒G型和P型的分型。方法 对马鞍山和苏州地区5岁以下急性腹泻儿童粪便标本1267份，采用酶联免疫吸附试验和巢式-聚合酶链式反应检测轮状病毒感染和血清型。结果 轮状病毒阳性标本378份，主要来自2岁以下婴幼儿；血清型G型是优势株共250株，其中G1100株，G28株，G3111株，G43株，G93株，混合感染4株，未能分型21株(8．40％)；马鞍山以G1型为主(58．54％)，苏州以G3型为优势株(47．85％)；P基因型两地区流行株为P[4]和P[8]，分别占52．33％和47．67％，G1P[4]成为仅次于G1P[8]的GP组合。结论 在苏州和马鞍山地区5岁以下儿童轮状病毒腹泻中，流行的血清型分别为G3和G1，P基因型流行株为P[4]和P[8]。     

上海地区婴幼儿腹泻的轮状病毒分子流行病学研究

目的 了解上海地区婴幼儿腹泻轮状病毒感染的分子流行病学特点。方法 收集复旦大学儿科医院1999年11月-2001年12月,住院的腹泻病儿童粪便标本1230份。采用聚丙烯酰胺凝胶电泳法检测轮状病毒基因组,用核酸杂交确定轮状病毒G血清型别。采用Kruskal-Wallis非参数统计法对不同G血清型别的临床变量作分析。结果 1230份标本中轮状病毒基因阳性493份(40．1％),其中RNA长型397份(80．5％),短型55份(11．2％),混合型18份(3．7％),未能分型23份(4．7％)。328份含有足量RNA的标本行RT-PCR扩增VP7全基因,254份标本(77．4％)获得阳性产物,进一步作杂交分型显示G1血清型141例(55．5％),占第一位;G3血清型70例(27．6％),占第二位;G2血清型24例(9．4％),占第三位;混合感染血清型16例(6．3％),占第四位;仅发现1例G4型(0．4％),未分型2例(0．8％)。所有G2型均为电泳短型,而G1、G3、G4均为电泳长型。四种型别轮状病毒感染患儿的平均发病年龄以及发热峰值、热程、每天腹泻峰次数、腹泻天数、呕吐天数差异无显著意义。结论 A组轮状病毒是目前上海地区腹泻儿童的主要致病原。在不同的3年中,轮状病毒的电泳型均以长型为主,流行血清型为G1-G3型,G4型很少见。未发现轮状病毒感染的临床特征与不同G血清型有关。     

5岁以下婴幼儿病毒性腹泻病原学分析和研究

目的 探讨甘肃白银市5岁以下婴幼儿病毒性腹泻感染状况、流行特点及轮状病毒(rotavirus,RV)血清型分型.方法 收集白银市第二人民医院2008年10月至2009年9月腹泻患儿粪便标本519份,其中住院181例,门诊338例,由中国疾病预防控制中心采用酶联免疫法进行RV、杯状病毒(human calicivirus,HuCV)、肠道腺病毒(enteric adenovirus,EADv)、人类星状病毒(human astrovirUS,HAstV)的检测,以及逆转录-聚合酶链反应法对RV检测阳性标本进行血清型分型.结果 检测的519份标本,4种病毒总阳性率为45.09％,其中RV 34.68％、HuCV 7.32％、EADv 2.12％、HAstV0.96％.RV是导致该地区婴幼儿病毒性腹泻的主要病原.180份RV阳性标本,G血清型流行优势菌株为G3型(35％),其次是G1型(20.6％),比例最低是G4型(2.2％).P基因型最常见的毒株类型是P8型(63.3％),其次是P4型(10％),最少的是P10型(0.6％).其发病高峰在每年10月、11月和12月为主,患病年龄以6～23月龄婴幼儿为主.结论 RV是导致白银地区儿童病毒性腹泻的主要病原,其次为HuCV、EADv及HAstV.RV亚型以[P8]G3为主,存在混合感染情况.     

兰州地区婴幼儿轮状病毒和杯状病毒腹泻的研究

目的 研究兰州地区婴幼儿轮状病毒和杯状病毒腹泻的分子流行病学特点。方法 2001年12月～2002年11月在兰州医学院第一附属医院儿科收集婴幼儿病毒性腹泻粪便标本244份，用酶联免疫吸附试验(ELISA)检测轮状病毒抗原，阳性标本用逆转录聚合酶链反应(RT-PCR)、巢式PCR及琼脂糖凝胶电泳方法进行基因分型；阴性标本用ELISA和RT-PCR进行杯状病毒病原检测。结果 224份标本轮状病毒检出率为45．5％(111／244份)，G分型中G3为58．6％，G1为18．0％，G2为4．5％，G9为2．7％，混合型为6、3％，11例未能分型。P分型中P[8]为61．0％，P[4]为7．3％，混合型为2．4％，14例未能分型。133份轮状病毒阴性标本杯状病毒检出率为15．8％，其中ELISA检测阳性率为11．3％，RT-PCR检测阳性率为9．0％。结论 轮状病毒是兰州地区婴幼儿病毒性腹泻的主要病原，2002年流行株为G3型，发现了G9型轮状病毒感染；杯状病毒也是引起兰州地区婴幼儿病毒性腹泻的病原之一，应引起重视。     

兰州地区婴幼儿轮状病毒腹泻的分子流行病学研究

目的通过分子流行病学调查研究兰州地区婴幼儿轮状病毒(RV)腹泻的分型特点。方法采用酶联免疫吸附试验(ELISA)和逆转录-聚合酶链反应(RT-PCR),对2003年7月~2004年6月兰州地区收集的624例婴幼儿腹泻粪便标本中的271份进行轮状病毒病原检测,并对阳性标本进行分型。结果ELISA法RV阳性率为56.46%(153/271),阳性率与性别无关。不同年龄段的感染率以6~11个月和12~23个月年龄段最高,分别为57.38%、63.33%,但各年龄段间无显著差异。不同月份的感染率具有显著性差异,发病高峰在10、11月份,分别为86.27%和73.81%。在ELISA阳性标本中RT-PCR法检出G3 94例(61.44%),G2 4例(2.61%),G9 3例(1.96%),未发现G1、G4型和混合感染,52例(33.99%)未能分型。G分型阳性标本中检出P[8]型28例(40.58%),其他未能分型。结论轮状病毒是兰州地区婴幼儿非细菌性腹泻的主要病原,其流行的主要血清型为G3型。     

2010年郑州地区婴幼儿腹泻A组轮状病毒VP7基因型分型

目的 研究2010年郑州地区秋冬季婴幼儿腹泻A组轮状病毒(RV)VP7基因型分型.方法 收集郑州市第一人民医院儿科2010年9-12月腹泻住院患儿的粪便标本80份.采用TRIzol法抽提粪便中RV RNA,采用聚丙烯凝胶电泳法检测其RV基因组,反转录(RT)-PCR法以及巢式(net)-PCR法进行A组RV的VP7基因型分型.结果 80份粪便标本中,40份检测到A组RV RNA基因,阳性率为50%.男24例,女16例;年龄6～36个月,平均11.7个月.对40份A组RV RNA阳性标本进行VP7基因型分型,其中G3型30例(75.0%),G1型6例(15.0%),G1和G3混合感染型3例(7.5%),G1与G9混合感染型1例(2.5%).40例A组RV RNA阳性患儿中并呼吸道感染7例,其中G3型6例,G1和G3混合型1例.结论 郑州地区秋冬季婴幼儿腹泻A组RV以G3型为主要流行基因型,并发现混合型感染病例及不常见的G9型感染病例.     

2009至2010年南京儿童医院5岁以下儿童病毒性腹泻分子流行病学特点

目的 对南京儿童医院（我院）5岁以下儿童轮状病毒(HRV)、人杯状病毒(HuCV)、星状病毒(AstV)和肠道腺病毒(AdV)感染的分子流行病学进行研究,为病毒性腹泻的防治提供基础数据和理论依据。 方法 收集2009年7月至2010年6月于我院消化科门诊就诊的5岁以下腹泻患儿的粪便标本。采用ELISA法检测A组HRV,阳性标本采用RT-PCR法进行毒株分型鉴定;HuCV、AstV和AdV采用RT-PCR或PCR法进行检测。 结果研究期间共收集病毒性腹泻患儿粪便标本300份,其中男188例,女112例。年龄1~59月龄,平均（10.5±9.2）月龄。4种病毒检测阳性率依次为HRV(37.7%,113/300)、HuCV(23.7%,71/300)、AstV(4.0%,12/300)和AdV(2.7%,8/300)。①HRV G血清型以G3型(38.9%)最常见,其次为G2型(8.8%);P基因型以P［8］（33.6%）为主。G血清型和P基因型组合以G3P［8］(15.9%)为主。113份HRV阳性标本中,7~12月龄儿童占43.4%（49/113）,高发季节为2009年10月至2010年1月。②HuCV感染中诺如病毒检出67份,扎如病毒4份。HuCV感染于2009年8月出现一个小高峰,发病年龄高峰为7~12月龄（38/71）,24月龄以下患儿占95.8%(68/71)。③10/12例AstV感染发生于2009年10月至2010年1月,月龄分布为2~16月龄。④8例AdV阳性标本PCR产物经克隆测序,AD 2、3、5、7、12和41型各1例,AD 31型2例。AdV感染高发于2010年1至6月,发病高峰为7~12月龄（4/8）。⑤60.0%（180/300）至少检出4种病毒中的一种。混合感染23份,其中12份（52.2%）为HRV+HuCV,5份（21.7%）为HRV+AstV,2份为HuCV+AstV, HRV+AdeV、HuCV+AdeV、AdeV+AstV、HRV+AstV+HuCV各1份。 结论 HRV是引起婴幼儿病毒性腹泻最主要的病毒病原,以G3P［8］为主要优势株。HuCV、AstV和AdV也是重要的病原。     

马鞍山、苏州地区婴幼儿轮状病毒腹泻临床特征和G血清型研究

目的　研究马鞍山和苏州地区婴幼儿轮状病毒 (RV)腹泻的临床特征和G血清型。方法　收集马鞍山和苏州地区5岁以下儿童急性腹泻粪便标本 1 2 67份 ,采用酶联免疫吸附试验和巢式 聚合酶链反应检测RV感染和血清型。结果　 1 2 67份标本中RV阳性 378份 ,血清型G型是优势株共 2 50株 ,其中G1 1 0 0株 ,G3 1 1 1株 ,G2 8株 ,G4、G9各 3株 ,混合感染 4株 ,未能分型2 1株 (8.40 % )。马鞍山地区以G1型为主 (58.54 % ) ,苏州地区以G3型为优势株 (47.85 % )。结论　RV是导致马鞍山和苏州地区婴幼儿腹泻的主要病原 ,流行的血清型分别为G1和G3型。     

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目的研究昆明地区2002~2004年轮状病毒肠炎患儿的病毒分子流行特征。方法收集昆明医学院第一附属医院2002、2003、2004年9~12月儿科住院和门诊轮状病毒肠炎患儿的粪便标本计210份。采用聚丙烯酰胺凝胶电泳法检测轮状病毒基因组,用逆转录-聚合酶链反应(RT-PCR)以及巢式-PCR(net-PCR)对5种主要的VP7的血清型(G1、G2、G3、G4和G9)进行分型,并采用RT-PCR对VP7阳性标本进行NSP4(非结构蛋白4)的基因扩增,选取30份标本用net-PCR法扩增出NSP4CDNA500bp送去测序。结果210份标本中轮状病毒基因阳性143份(68·1%),其中长型143份,未发现短型和混合型。143份阳性标本行RT-PCR扩增VP7全基因,134份获得阳性产物,进一步用net-PCR分型显示G3型73例(73/134,54·5%),G1型38例(28·4%),未分型11例(8·2%),混合型8例(6%),G4型4例(3%),未发现G2和G9型。所有VP7阳性标本均可以扩增出NSP4的全长,其中30份标本送去测序显示有变异。结论A组轮状病毒是目前昆明地区婴幼儿腹泻的主要致病原,在不同的3年中,轮状病毒电泳型以长型为主,血清型以G3、G1型为主,G4型少见,未见G2、G9型。3年监测结果显示NSP4存在变异,但未发现轮状病毒肠炎临床症状与NSP4的变异有关。     

苏州地区5岁以下儿童轮状病毒腹泻的临床流行病学研究

目的　 了解苏州地区 5岁以下儿童轮状病毒腹泻的临床、分子流行病学特征及其影响因素。 方法 　对 2 0 0 1年 9月— 2 0 0 3年 6月在苏州大学附属儿童医院就诊的 5岁以下腹泻患儿进行调查 ,收集粪便标本 ,检测轮状病毒 ,对阳性标本用ELISA、PCR法进行G、P分型 ;同时填写调查表 ,分析流行特征。资料用SPSS 10 0统计软件进行分析。 结果 　调查 9412例患儿 ,收集标本 2 0 48份 ,检测标本 13 41份 ,轮状病毒阳性 5 16份 ,阳性率 3 8 48% ;血清分型发现 ,G型以G3 ( 62 44 % )和G1( 2 4 41% )为主要流行株 ,P分型以P4( 5 8 0 2 % )为主 ;2 0 0 1年 11月— 2 0 0 2年 3月及 2 0 0 2年 10月— 2 0 0 3年 2月出现两个轮状病毒高峰 ;患者最小年龄为 12d ,3～ 2 3月龄占 90 12 % ,男女比例为 1 98∶1;1岁以内母乳喂养儿比人工及混合喂养儿阳性率均低 (P <0 0 1) ;轮状病毒腹泻多引起脱水。 结论 　苏州地区 5岁以下儿童腹泻以轮状病毒为主要病原体 ,其中G3型和P4型为主要流行株 ;秋冬季节为发病高峰 ;3～ 2 3月龄为高发人群 ;母乳喂养儿阳性率较低 ;轮状病毒腹泻多会引起脱水。     

Molecular epidemic features and variation of rotavirus among children with diarrhea in Lanzhou,China, 2001–2006

Background  Human rotavirus (HRV) is the most common pathogen causing severe diarrhea among infants and young children worldwide. This study aims to understand rotavirus epidemiology and its variation in the period of 2001–2006 in Lanzhou, Gansu Province, China, and to provide an epidemiological basis for the development of rotavirus vaccine. Methods  A total of 1019 stool specimens were collected from patients with acute diarrhea admitted to the First Hospital of Lanzhou University from 2001 to 2006, who were younger than 5 years old. Dako IDEIATM kits were used for detection of rotavirus, and RT-PCR was performed for determination of G serotype and P genotype of the rotavirus. Results  Rotavirus was present in 51.6% (526) of the 1019 specimens. G serotype identified G3 at 40.9%, G2 14.6%, G1 22.2% and G9 1.9%. Mixed-G infection was observed in 4.4% and non-typeable infection 16.0%. P genotype was observed in 372 samples, of which P[8] accounted for 186 cases (50.0%), P[4] 72 cases (19.4%), mixed-P infection 2 cases (0.5%), and non-typeable cases 112 (30.1%). G3 was the most prevalent G serotype found in this study from 2001 to 2004, G2 was the most prevalent G serotype (34.4%) from 2004 to 2005, and G1 (61.5%) was the most prevalent strain from 2005 to 2006. G9 was detected in 10 cases (1.9%) and G4 was not detected during this 5-year period. P[8] was the most prevalent P genotype found over the 5 consecutive years of this study, although there was a significant transition of P genotype from 2004 to 2005 with P[4] (45%) identified as the predominant P genotype, followed by P[8] (22.1%). The predominant G-P combination was P[8]G1 (33.6%), followed by P[8]G3 (32.1%) and P[4]G2 (17.2%). Rotavirus diarrhea admissions peaked between October and December. Continuous surveillance showed that the incidence rate of rotavirus was the highest in infants aged 6–23 months, averaging 11.0–11.9 months. Conclusions  Five years of continuous surveillance showed that rotavirus remains the most significant viral agent causing diarrhea hospitalization among children under 5 years old in Lanzhou, China although the predominant strain of rotavirus varies between years. Mixed-G serotype infection also appears to occur at a relatively high rate in Lanzhou.     

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目的通过分子流行病学调查研究兰州地区婴幼儿病毒性腹泻的病原学特点。方法采用酶联免疫吸附试验(ELISA)及逆转录聚合酶链反应(RTPCR),对2003年7月至2004年6月兰州地区收集的624例婴幼儿腹泻粪便标本随机抽取271例进行轮状病毒(RV)、杯状病毒(HuCV)及星状病毒(AstV)检测。结果在271例标本中共检出RV感染153例(56.46%),其中G394例(61.44%),G24例(2.61%),G93例(1.96%),未发现G1、G4型和混合感染;在随机抽取的69例G分型阳性标本中,检出P[8]型28例(40.58%);RV的感染对象主要为6～23月龄的婴幼儿,发病高峰在10、11月份(86.27%、73.81%)。在118例RVELISA阴性标本中检出HuCV感染13例(11.02%),其中诺如病毒(NLV)GⅡ型11例,札如病毒(SLV)2例,未检出NLVGⅡ型,发病年龄1～18个月(11.31±4.53个月);同时检出AstV感染7例(5.93%),发病年龄4～12个月(8.27±2.69)个月,其中有1例合并有SLV感染,另有1例为迁延性腹泻。HuCV和AstV感染均未表现出明显的季节性。结论RV是兰州婴幼儿病毒性腹泻的主要病原,其流行的主要血清型为G3型,HuCV和AstV亦是重要病原之一。     

Overview of rotavirus infections in Korea

Rotavirus is the most common cause of acute watery diarrhea in young Korean children. Rotavirus vaccine will soon be available, and information is urgently required about the serotype distribution of recent epidemics and clinical characteristics of rotavirus infection in Korean children before the implementation of a vaccination program against rotavirus. We reviewed published studies of the past 20 years, carried out on Korean children with rotavirus gastroenteritis. Rotavirus was estimated to be responsible for 46% of 4668 hospitalized Korean children with acute gastroenteritis. Rotavirus gastroenteritis was most prevalent among children aged 6-24 months, which accounted for 84% of all cases. Asymptomatic rotavirus infection was common. Rotavirus was one of the most commonly identified enteric pathogens in nosocomial diarrhea. Vomiting, respiratory symptoms and fever were prominent symptoms in rotavirus gastroenteritis. Transient elevation of liver enzymes, pulmonic infiltration and rarely afebrile convulsion were also observed. The epidemic peak, which occurred in November of the last 15 years, has been moving toward late winter and early spring in recent years. No apparent cause has been found to explain this alteration of peak seasonality. All serotyping studies in Korea for the past 10 years until 1997 revealed that G1 was most prevalent (45-81%). Interestingly, the predominant G serotype of the recent outbreaks in 1998 and 1999 was not G1 but G4. Approximately 95% of rotavirus isolates in recent outbreaks belonged to serotype G1, 2, 3 or 4.     

Characterization of rotavirus infection in a hospital neonatal unit in Pretoria,South Africa

Rotavirus infection in neonates is common and has been reported to be generally asymptomatic. In this longitudinal study, specimens were collected from 114 newborns in the Neonatal Unit at Pretoria Academic Hospital on a daily basis between January and May 1997. The babies remained in the ward between 1 week and 4 months. The stool specimens or rectal swabs were analysed for the presence of rotavirus antigen using a commercial enzyme-linked immunosorbent assay (Dako Rotavirus EIA) or electron microscopy. In total, 80 (70 per cent) of the neonates excreted rotavirus during their stay in the unit. There was a direct correlation between the length of stay in the ward and the shedding of rotavirus. The babies excreted rotavirus on average between 2 and 7 days. Rotavirus infection tended to occur within the first 2 weeks of life and was only observed once in most babies. Polyacrylamide gel electrophoresis of the RNA revealed the presence of two strains of rotavirus, with the differences in the RNA electropherotype occurring in the RNA segment triplet 7, 8 and 9. The VP7 serotype of the virus is encoded by one of these genes, and so the VP7 serotype of the virus was determined by monoclonal antibody and RT-PCR using VP7 serotype specific primers. The VP4 genotype of the viruses was also determined using RT-PCR of the VP4 gene to determine if a new rotavirus had been introduced to the ward. The strains were all characterized as G4P[6], which is similar to the antigenic make-up of the virus recovered 10 years before. This highlights the remarkable stability of rotavirus strains in neonatal units over long periods of time.     

改进探针标记法斑点杂交在轮状病毒VP7分型中的应用

目的建立一种更加简便的A组人轮状病毒（HRV）核酸斑点杂交VP7分型方法,以便用于对HRV流行情况进行调查。方法在HRV VP7编码基因各G基因型间高度变异而型内高度保守区域设计分型探针,在该区域的两侧相对保守区域设计一对通用引物,利用PCR分别将地高辛标记HRV 5种常见型别（G1～4,G9型）的DNA探针,建立基于VP7的斑点杂交方法;选取经抗原检测和聚丙烯酰胺凝胶电泳（PAGE）检测均为HRV阳性的2006至2008年住院腹泻患儿粪便标本200份,RT-PCR扩增VP7全基因,并对扩增阳性产物应用斑点杂交方法进行G型别分析。结果建立的斑点杂交方法在5种型别探针间无交叉反应,各型探针的检测灵敏度可达到10 pg。200份PAGE阳性标本中162份RT-RCR扩增VP7基因阳性,斑点杂交显示G1型41例(25.3%),G2型2例(1.2%),G3型63例(38.9 %),G9型35例(21.6%),混合感染19例（11.7%）,杂交未分出型2例(1.2%),未检测到G4型HRV。结论本研究所建立的斑点杂交方法敏感度和特异度强,适合在HRV大规模分子流行病学调查时应用。通过该方法的初步应用,发现北京地区婴幼儿HRV除了常见的G1、G2和G3型外,还有G9型感染。     

VP6 subgroup and VP7 serotype of human rotavirus in Zaria, northern Nigeria

A survey of rotavirus infection in infants and young children with acute diarrhoea was undertaken in Zaria, northern Nigeria during 1997 and 1998. In total, 375 faecal specimens were collected from children aged between 1 and 60 months and 122 specimens from age-matched control children without diarrhoea. Fourteen specimens were collected from neonates in the University Teaching Hospital. Rotavirus antigen was detected in 61 diarrhoeal and four control specimens; four neonates were shedding rotavirus. Polyacrylamide gel electrophoresis of the viral genome showed the presence of five strains of rotavirus with long RNA electropherotypes and one short pattern. The rotavirus VP6 subgroup was determined by monoclonal antibodies specific ELISA and showed that subgroup II strains predominated (72 vs. 9.8 per cent), while eight strains could not be subgrouped and three did not react at all. Examination of the VP7 serotype showed G1 and G3 strains circulating at similar levels (29 and 25 per cent), but no serotype G2 nor G4 strains were identified. G1/G3 'mosaic' virus strains circulated commonly (10 per cent).     

Molecular Epidemiology of Rotavirus Strains in Under Five Children

Objectives

To determine the prevalence and genotype distribution of rotaviruses in children presenting to authors’ hospital in Middle Black Sea region of Turkey. The results may supply important information about vaccine studies in Turkey.

Methods

Rotavirus antigen was detected by latex agglutination test and rotavirus RNA was detected by RT-PCR test. On the other hand, rotavirus positive samples were genotyped by semi-nested multiplex polymerase chain reaction.

Results

The highest rate of rotavirus positivity (46.9%) was observed among children in the 13 to 24 mo age group. All the positive-strains were in G1–4, G8–9, P [4], P [8], and P [9] genotypes. The most common G and P combination in present study was G9P[8] (n = 24, 28.9%).

Conclusions

The present results indicated that the most prevalent genotypes were G1, G9, P8, G9P[8] and G1P[8] in authors’ region. Rotavirus vaccines used in this region must include mainly these genotypes.