秋冬季婴幼儿轮状病毒感染分子流行病学调查

目的监测2010年云南昭通地区秋冬季腹泻患儿感染轮状病毒的流行趋势,了解流行毒株的基因型特征。方法 2010年9—12月在云南省昭通市第一人民医院采集94份就诊的腹泻患儿粪便,应用胶体金法、逆转录聚合酶链式反应及PAGE电泳等方法,对样品进行轮状病毒抗原检测,其中8份阳性样品进行VP7基因测序分析及基因型分型。结果腹泻病患排泄物中轮状病毒抗原阳性比例为54.3%(51/94);8份轮状病毒基因序列分析,其中7份为G1型,1份为G3型;核苷酸同源性分析发现检测样品中轮状病毒主要与在中国辽宁、印度和泰国发现的流行株相似。结论 2010年昭通地区秋冬季患儿腹泻主要由轮状病毒引起,其中以G1型为主,其次为G3型。     

北京地区2007-2008年G9型A组人轮状病毒VP7和VP4基因分析

目的 了解北京地区2007-2008年检测到的G9型A组人轮状病毒外壳蛋白VP7和VP4的基因特征.方法 选取经过轮状病毒核酸杂交方法检测为G9型轮状病毒的12份儿童腹泻患儿的粪便标本,应用针对VP7全长基因的特异引物对进行RT-PCR扩增,对所获得的VP7全长基因进行克隆和测序,将所获得的序列与GenBank中的G9型原型病毒株和近期流行株的VP7基因进行序列和种系进化分析;经巢式PCR对G9型的VP4进行P基因分型.结果 12株G9型轮状病毒经VP7基因的序列比较分析得到确认.P基因分型结果显示北京地区近年来存在G9P[8]和G9P[6]型两种组合的轮状病毒感染.序列和种系进化分析发现北京G9型株VP7基因与世界范围内近期流行的G9型株一样都属于进化分支Ⅲ,彼此间的核苷酸和氨基酸同源性较高,而与国内最早报道的G9型T203进化关系较远,且北京G9P[8]和G9P[6]型株分别与国内近期报道的新疆G9P[8]和G9P[6]型株及相应的武汉G9型株VP7基因,在氨基酸位点上存在一些共同的氨基酸残基取代.结论 北京地区近年存在G9P[8]和G9P[6]两种不同基因组合的G9型轮状病毒感染,需要进一步加强对G9型轮状病毒的分子流行病学监测.     

福州地区2009-2014年腹泻儿童轮状病毒特征

目的 了解轮状病毒在福州地区腹泻儿童中的流行情况.方法 收集2009-2014年5岁以下腹泻住院儿童粪便标本,用ELISA法检测轮状病毒抗原,RT-PCR法确定基因型别.对G9轮状病毒阳性标本的VP7基因全长测序及进化分析.结果 福州地区腹泻儿童轮状病毒高峰期在10～12月,呈单峰流行态势.G9轮状病毒在2011年后成为福州地区优势流行型别,毒株VP7基因核苷酸序列相似性92.5％～100％,毒株间有较高的同源性,进化分析都属于G9第3亚型.结论 G9轮状病毒在福州地区流行强度逐渐加强,目前已成为优势流行型别.毒株同源性高,属G9型3亚型.     

宁波市一起新生儿轮状病毒腹泻爆发的调查

目的 了解宁波市轮状病毒(Rotavirus,RV)地方株的基因型,为中国轮状病毒疫苗的研制提供资料.方法 通过逆转录-聚合酶链反应获得宁波市RV流行株VP7基因全序列并进行序列测定.结果 Ningbo 06-1和Ningbo 06-3 VP7氨基酸同源性为90.2%,且均与G1型的标准株Wa同源性较高,分别为95.1%和87.7%,而与其它血清型代表株同源性均<85%.在VR5和VR8两个高变区域,Ningbo 06-1和Ningbo 06-3与Wa的同源性分别为89.3%和75.0%,与其它血清型代表株同源性分别低于67.9%和60.7%.结论 宁波市A组RV以G1血清型第一亚组为主.     

青海省67份轮状病毒结构蛋白VP7基因分型和分布情况

目的 研究2016-2017年青海地区腹泻患者轮状病毒基因分型及流行病学分布。方法 收集门诊及住院部腹泻的粪便标本238份,采用实时荧光PCR法对轮状病毒A组进行检测,阳性标本进行VP7基因扩增和测序。结果 238份粪便标本中,通过实时荧光PCR 检测到轮状病毒A组阳性67份,阳性率为28.15%（67/238）;对67份轮状病毒A阳性标本进行VP7蛋白检测和测序,测序后得到29份核苷酸序列,用Clustral X Bootstap NJ Tree软件构件进化树,分析发现2016年3月-2017年12月青海轮状病毒以G9P8型为主,共26株,占89.66% (26/29),G2P4型2株(2/28),G3P8型1株(1/28), 轮状病毒腹泻发病高发季节为9-12月,其中以12月份检出最高,占总数的61.19%（41/67）。病人以成人为主,成人和5岁以下儿童比例为1.73[DK]∶1。结论 2016-2017年青海地区轮状病毒以流行病毒株G9P8型为主。     

辽宁省首次检出G9型A组轮状病毒

目的了解辽宁省A组轮状病毒的感染情况,为轮状病毒的预防控制提供科学依据。方法采集辽宁省沈阳、大连、丹东、阜新4个市门诊及住院疑似病毒性腹泻患者粪便标本135份,采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)扩增A组轮状病毒VP7基因和VP4基因,RT-PCR产物进行核苷酸碱基序列的测定和比对,并构建VP7基因遗传进化树。结果 135份粪便标本中,共检测出A组轮状病毒阳性标本21份;A组轮状病毒G基因分型:G9型15株,G3型2株,G2型1株,G1型1株,未分型2株;P基因型分型:P[8]型20株,P[4]型1株;G/P基因型组合以G9P[8]为主共15株,G3P[8]2株,G1P[8]1株,G2P[4]1株,G/P[8]2株。结论辽宁省首次检出G9型A组轮状病毒,主要流行G/P基因型组合为G9P[8]。     

中国1998～2004年G9型轮状病毒分子流行病学研究

目的研究中国流行的轮状病毒(Rotavirus,RV)G9型毒株的分子流行病学特征。方法在中国9个地区收集5岁以下腹泻患儿粪便标本,应用酶联免疫吸附试验(ELISA)方法检测RV,对阳性标本用逆转录-聚合酶链反应(RT-PCR)分型,选择G9型毒株进行VP7基因全长克隆测序和分子流行病学分析。结果1998~2004年共检测出RV1 268份,其中45份为G9型(3.5%),昆明最多(34/45),其次是兰州(8/45)、长春(2/45)、卢龙(1/45),北京、郑州、杭州、福州、广州未检测到G9型毒株。对35份G9型标本进行P分型鉴定:15份为P[8]型,12份为P[6]型,5份为P[4]型,1份为P[8 4]混合型,2份未能分型。中国1998~2000年以P[8]G9毒株流行为主,而2001年后以P[6]G9毒株为主。22株G9型病毒VP7基因序列比对结果表明,中国流行的G9型毒株彼此同源性高,同属G9第3亚型。结论中国流行的RV G9型株与世界各地的流行株同源性较高,国内传播范围扩大的趋势值得关注。     

A组轮状病毒G3型哈尔滨地方株VP7基因序列分析

目的:了解我国A组轮状病毒G3型哈尔滨地方株与标准株及国内外部分地区G3型地方株VP7蛋白基因序列的差异,为轮状病毒疫苗在我国东北地区的研发与应用提供资料。方法:通过一步法RT-PCR获得了6株轮状病毒哈尔滨地方株VP7蛋白的cDNA片段,,对其进行扩增、克隆、测序,用DNASTAR生物软件进行序列分析。结果:6株哈尔滨地方株VP7蛋白推导氨基酸序列同源性99.4%～100%;与G3标准株Crw-8的同源性为94.2%;与G1、G2、G4标准株比对变异较大(75.2%～82.2%);哈尔滨地方株氨基酸序列在aa108、aa266、aa268、aa278位点的变异完全相同。结论:6株轮状病毒G3型哈尔滨地方株来源于相同的一支G3型轮状病毒毒株。     

2009年河南肠道病毒71型VP1基因特征分析

目的了解河南省2009年肠道病毒71型（EV71）流行株VP1基因特征。方法采用RT-PCR方法从手足口病患者粪便、咽拭子样品中扩增VP1区全长基因和序列测定,利用生物信息学软件对序列进行分析,构建序列遗传发育树。结果 262份样品中,VP1基因RT-PCR扩增鉴定结果显示111份样品阳性,选取其中20份样品进行VP1基因全序列测定,结果显示其与C4亚型代表株核苷酸同源性为90.8%～93.6%,氨基酸同源性为94.0%～99.3%。结论河南省手足口病患者感染EV71病毒的流行株属C基因型的C4亚型。     

河南省轮状病毒VP7基因型的分布

1990-1996年间由河南五市县5岁以下急性胃肠炎患儿获得的188份轮状病毒RNA电泳阳性粪便标本,经G血清型(VP7)特异1、2、3和4型寡核苷酸探针枪查,132份(70%)与1、2或3型探针杂交,其中60%为G血清1型,27%为G血清2型,11%为G血清3型,有3份标本为G_1+G_3混合型(2%),未检测到G血清4型。同时发现2株电泳短型,1株电泳长型,分别与G1型和G2型探针杂交。     

重庆地区婴幼儿轮状病毒腹泻VP7型别分析

目的：研究重庆地区1998－2000年度秋冬季婴幼儿轮状病毒腹泻分子流行病学,方法：采用逆转录－聚合酶链反应（RT－PCR）扩增婴幼儿腹泻便样中的编码轮状病毒VP7蛋白的全基因片段（1062bp),再用巢式－聚合酶链反应（net-PCR)对扩增得到的VP7基因进行分型,同时利用核苷酸序列分析方法进行分型。结果：在1998－1999年度130例婴幼儿腹泻便样中VP7基因阳性者50例（38．46％）,其中G1型占88％（44／50）,G3型占8％（4／50）,混合型占4％（2／50）,均为G1＋G3型,而1999－2000年度轮状病毒流行季节采集的112 标本中VP7基因扩增阳性者38例（33．93％）,其中G3型占78．95％（30／38）,G1型占13．16％（5／38）,混合型占7．89％（3／38）,均为G1＋G3型,苷酸序列分型结果与PCR分型结果一致。结论：重庆地区 1998－1999年度轮状病毒流行季节中流行的轮状病毒以G1型为主,而1999－2000年度轮状病毒流行季节中G3型为主,在连续两年的监测中出现轮状病毒血清型的转变。     

Clinical and genetic characteristics of unusual G12P[11] rotavirus strains recovered from neonates: A study from Pune,Western India

Rotavirus infections in neonates are generally nosocomial, and differ from pediatric infections both clinically and epidemiologically. These infections are predominantly asymptomatic and often associated with unusual strains. Globally, so far limited data is available on rotavirus infections in neonates admitted at Neonatal Intensive Care Unit. The aim of the present study is to determine the prevalence of rotavirus among neonates and to study their genetic characteristics. Stool specimens (n = 701) collected from neonates (n = 621) admitted during April 2016 to March 2018 mainly for prematurity, low birth weight and associated respiratory distress syndrome from two hospitals from Pune were tested for rotavirus, genotyped and representative strains were sequenced for the genes encoding outer capsid proteins, VP7 and VP4. Rotavirus was detected in 24.31% neonates. Majority of rotavirus infected neonates (98.68%) were asymptomatic. Peak rotavirus antigen detection (91.38%) occurred during the first 2 weeks of admission. Low, very low and normal birth weight neonates with gestational age ≥28 weeks had significantly higher rotavirus infection than those with extreme low birth weight with gestational age <28 weeks. Rotaviral infections occurred almost evenly throughout the year without an apparent peak in colder months. Predominance of unusual G12P[11] strains (97.1%) was observed. Phylogenetic analysis of the partial VP7 coding gene revealed all G12 strains clustered in lineage III and shared 96.94%–100% (nucleotide) and 96.26%–100% (amino acid) identities among themselves, and 95.69%–98.98% (nucleotide) and 94.77%–98.98% (amino acid) with other lineage III G12 strains respectively. Similarly VP4 partial gene sequences of P[11] study strains shared 97.5%–100% (nucleotide and amino acid) identities among themselves and highest 93.34%–94.53% (nucleotide) and 93.57%–94.64% (amino acid) identity with vaccine strain 116E, G9P[11]. The study highlights high frequency of unusual G12P[11] strains among neonates for the first time in western India and reaffirms limited strain diversity in this population. The knowledge of neonatal strains is important for estimating the efficacies of rotavirus vaccines.     

Rotavirus strains circulating in Africa during 1996-1999: emergence of G9 strains and P[6] strains

Rotavirus infection is associated with 150000-200000 deaths annually in Africa. Although the withdrawal of the RotaShield vaccine has been a major setback in rotavirus vaccine development, new vaccine candidates are under development and approaching phase II and III trials. Before these trials could be conducted in Africa, a comprehensive survey of the circulating VP7 serotypes and VP4 genotypes is required. During the past 3 years, over 3000 rotavirus-positive specimens from several African countries have been analysed. RT-PCR techniques for the VP7 and VP4 genotypes and by monoclonal antibodies to the VP6 subgroup and VP7 serotype have been performed. Almost 75% of the strains were typed by the VP7 monoclonal antibodies or RT-PCR. VP4 genotyping was done in approximately half of these strains. The predominant strains circulating across Africa during 1996-1999 were P[6]G1 and P[6]G3 strains. Geographic differences were noted and West Africa displayed the most diverse strains with G3/8 and G1/3 "mosaic" viruses occurring commonly. G9 strains were identified in several countries indicating that the strain is emerging in Africa too. G9 was the predominant strain in certain countries during 1999. The circulating types observed will have implications for the new rotavirus vaccine candidates.     

山东省1990 ～ 1998 年肾综合征出血热流行特征及其变化趋势

山东省是我国肾综合征出血热 (ＨＦＲＳ)流行最严重的疫区 ,其发病人数占全国发病总数的 1/3左右[1] 。笔者分析了山东省ＨＦＲＳ流行特征及其变化 ,旨对ＨＦＲＳ综合防制工作有一定的指导作用1 .流行概况 :在 1995～ 1998年ＨＦＲＳ发病率为 19.19/10万 ,病死率为 0 .42 % ,分别比 1990～ 1994年发病率 10 .15 /10万和病死率 0 .90 %上升和下降 (χ2 =12 784.47,Ｐ <0 .0 0 0 1;χ2 =10 5 .12 ,Ｐ <0 .0 0 0 1)。ＨＦＲＳ发病高峰在 1995年 ,发病率为 2 6 .0 8/10万 ,病死率为 0 .48%。山东省共有141个县 (区 ) ,1990～ 1998年ＨＦＲＳ发病…     

Porcine rotavirus strain Gottfried-based human rotavirus candidate vaccines: construction and characterization

Rotavirus gastroenteritis remains the leading cause of severe diarrheal disease in infants and young children worldwide, and thus, a safe and effective rotavirus vaccine is urgently needed in both developing and developed countries. Various candidate rotavirus vaccines that were developed by us and others have been or are being evaluated in different populations in various parts of the world. We have recently confirmed that a porcine rotavirus Gottfried strain bears a P (VP4) serotype (P2B[6]) closely related to human rotavirus P serotype 2A[6] which is of epidemiologic importance in some regions of the world. Based on the modified Jennerian approach to immunization, we have constructed 11 Gottfried-based single VP7 or VP4 gene substitution reassortant vaccine candidates which could provide: (i) an attenuation phenotype of a porcine rotavirus in humans; and (ii) antigenic coverage for G serotypes 1-6 and 8-10 and P serotype 1A[8], 1B[4] and 2A[6]. In addition, following immunization of guinea pigs with Gottfried VP4, we found low but consistent levels of neutralizing antibodies to VP4 with P1A[8] or P1B[4] specificity, both of which are of global epidemiologic importance. Thus, porcine-based VP7 reassortant rotavirus vaccines may provide an advantage over rhesus- or bovine-based VP7 reassortant vaccines since the VP4s of the latter vaccines do not evoke antibodies capable of neutralizing the viruses bearing P1A[8], P1B[4] or P2A[6] VP4.     

昆明地区22株人轮状病毒VP7及NSP4基因分析

目的研究昆明地区轮状病毒（RV）不同VP7血清型及NSP4基因变异与腹泻的流行及症状严重程度的关系。方法对2002年和2003年分离于昆明地区的RV，用PCR分型法对四种主要的VP7血清型进行分型，并对从150份RV腹泻标本VP7血清型为G1、G3、（34型RV株中挑出的14份一般腹泻株和8份重症腹泻株的NSP4基因序列进行了分析，与来自GenBank Database的4株人RV（Wa、KUN、AU-1、Hochi）和3株动物RV（EW、OSU、SA11）以及中国不同地区流行株NSP4的基因变异情况进行了比较。结果2002年昆明地区RV流行株以G1型为主，2003年RV腹泻株以G3型为主；昆明地区RV流行株间的氨基酸同源性高达98．9％～99．4％，22株流行株全都属于Wa组；NSP4变异与地域及VP7血清型无关；NSP4基因变异与RV腹泻临床症状严重程度不相关（P〉0．05）。结论不同年份相同季节不同地域流行的RVVP7血清型变异较大，而NSP4基因的相对保守性及其免疫原性使其有可能成为发展疫苗的候选基因。     

Rotavirus strain distribution in Ghana pre- and post- rotavirus vaccine introduction

BackgroundGhana introduced the monovalent rotavirus vaccine (Rotarix) into its national paediatric vaccination programme in May2012. Vaccine introduction was initiated nationwide and achieved >85% coverage within a few months. Rotavirus strain distribution pre- and post-RV vaccine introduction is reported.MethodsStool samples were collected from diarrhoeic children <5 years of age hospitalized between 2009 and 2016 at sentinel sites across Ghana and analyzed for the presence of group A rotavirus by enzyme immunoassay. Rotavirus strains were characterized by RT-PCR and sequencing.ResultsA total of 1363 rotavirus EIA-positive samples were subjected to molecular characterization. These were made up of 823 (60.4%) and 540 (39.6%) samples from the pre- and post-vaccine periods respectively. Rotavirus VP7 genotypes G1, G2 and G3, and VP4 genotypes P[6] and P[8] constituted more than 65% of circulating G and P types in the pre–vaccine period. The common strains detected were G1P[8] (20%), G3P[6] (9.2%) and G2P[6] (4.9%).During the post-vaccine period, G12, G1 and G10 genotypes, constituted more than 65% of the VP7 genotypes whilst P[6] and P[8] made up more than 75% of the VP4 genotypes. The predominant circulating strains were G12P[8] (26%), G10P[6] (10%) G3P[6] (8.1%) and G1P[8] (8.0%). We also observed the emergence of the unusual rotavirus strain G9P[4] during this period.ConclusionRotavirus G1P[8], the major strain in circulation during the pre-vaccination era, was replaced by G12P[8] as the most predominant strain after vaccine introduction. This strain replacement could be temporary and unrelated to vaccine introduction since an increase in G12 was observed in countries yet to introduce the rotavirus vaccine in West Africa. A continuous surveillance programme in the post-vaccine era is necessary for the monitoring of circulating rotavirus strains and the detection of unusual/emerging genotypes.     

昆明市儿童医院1998～2001年轮状病毒哨点监测分析

目的 了解昆明市轮状病毒腹泻的流行状况。方法 以昆明市儿童医院为哨点监测,监测对象为5岁以下腹泻住院患儿,收集患儿的临床资料和粪便标本进行轮状病毒的检测和分型。病毒检测用聚丙烯酰胺凝胶电泳(PAGE)和酶联免疫吸附试验(ELISA),毒株分型用ELISA和/或反转录-聚合酶链反应(RT-PCR)。结果 3年监测中共收集466份腹泻患儿的粪便标本,轮状病毒的检出率为52.8％(246/466)。轮状病毒感染97％发生于2岁以下儿童。感染有明显的季节性,10～12月份是流行季节。对204份轮状病毒阳性标本进行G分型,G1型为流行优势株,占47.5％,其次为G2型(17.6％)、G3型(15.7％)G9型(4.9％)和G4型(1.0％)。P基因型以P[4]、P[8]和P[6]型为常见。最常见的P-G组合型是P[4]G2,占34.1％(14/41),其次是P[8]G1和P[6]G9,分别占29.3％(12/41)和12.2％(5/41),还有其他7种不常见的P-G组合的毒株类型。结论 轮状病毒是昆明地区儿童腹泻住院的主要病原,毒株呈现型的多样性,应该开发和应用轮状病毒疫苗预防控制其流行。     

Rotavirus serotype G9P[8] and acute gastroenteritis outbreak in children, Northern Australia

During 2001, an outbreak of severe acute gastroenteritis swept through Central and northern Australia and caused serious disruption to health services. We tracked and characterized the rotavirus strain implicated in the outbreak. Comparison of the electropherotypes of outbreak samples suggested that one G9P[8] strain was likely responsible for the outbreak. Samples were obtained from geographically distinct regions of Australia where the epidemic had occurred. The outbreak strains showed identical nucleotide sequences in genes encoding three rotavirus proteins, VP7, VP8, and NSP4, but they were distinct from G9P[8] strains isolated in previous years. Several of the amino acid substitutions on the VP7 and NSP4 proteins were identified in regions known to influence function and may have contributed to the emergence and increased dominance of the outbreak strains. Rotavirus serotype surveillance should continue with methods capable of identifying new and emerging types.