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。     

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。     

不数民族新生儿及婴幼儿中人轮状病毒分子流行病学研究

用降丙烯酰胺凝胶电泳技术,对内蒙古自治区三个不同地理位置的少数民族自治旗及一个城市汉族对照区秋冬季采集的新生儿胎便及婴幼儿急性腹泻便进行轮状病毒ＲＮＡ基因组特点分析。共检测胎便标本１４６份,腹泻便标本１１６份。结果表明;几个少数民族地区流行的轮状病毒均以Ａ组Ⅱ亚组为主,并在一个地区有多种差异电泳型存在,新生儿胎便中可检出轮状病毒并显示了特征性Ａ组轮状病毒ＲＮＡ基因型。胎便中轮状病毒检出率高的地区,     

哈尔滨地区婴幼儿腹泻轮状病毒优势株的变化

本文对1988—1989年哈尔滨地区婴幼儿腹泻进行了轮状病毒核酸电泳型调查,并于1984—1985年资料比较,以探明这一地区轮状病毒流行优势株的变化。 标本:婴幼儿腹泻粪便标本采自哈尔滨医科大学附属二院儿科及哈尔滨红十字儿童医院。病毒RNA提取:参考Herring方法。聚丙烯酰胺电泳(PAGE):参考Uaemmli方法。 结果:123份急性婴幼儿腹泻粪便标本PAGE检测,47例呈现轮状病毒RNA图型,阳性率为38.2％。电泳图型的基本模式为4:2:3:2,表明均为A组轮状病毒。根据各片段的迁移位置差异,共可见9种不同的电泳型,其中短型3例,占6.4％,含2个电泳型;长型44例,     

腹泻患儿粪便A群轮状病毒抗原检测的结果分析

目的:了解本地区腹泻婴幼儿的A群轮状病毒感染情况及其流行特点。方法:采用胶体金法对我院2009年10月-2010年9月2104例有腹泻和肠炎特征的婴幼儿粪便进行A群轮状病毒抗原检测。结果:在2104例受检者中,A群轮状病毒感染的总阳性率是24.71%,其中男性感染率24.17%,女性为25.40%。不同年龄组间以1-2岁婴幼儿感染率最高,为32.13%,0-1岁为20.72%,2-5岁为12.03%。感染的季节特征是秋末冬初季(10-12月)阳性率最高,为42.82%,春末夏初季(4-6月)最低,为8.81%。结论:由A群轮状病毒感染引发的急性腹泻主要发生在1-2岁的婴幼儿,各个季节均有发生,以秋末冬初季为高发。     

轮状病毒感染相关受体的研究进展

轮状病毒是引起婴幼儿腹泻的主要病原体之一,由三层同心的蛋白质衣壳包被双链RNA基因组构成。轮状病毒感染宿主细胞主要依赖于病毒识别细胞表面的特异性受体并与其发生结合,因此,受体是病毒感染细胞的重要因素。目前已发现的受体包括唾液酸、整合素、Toll样受体、血型组织抗原等,本文将对参与轮状病毒感染的相关受体作一简要概述。     

苏浙闽地区婴幼儿急性腹泻患者病毒检测及 临床特征分析

目的分析苏浙闽地区婴幼儿急性腹泻患者常见病毒及患儿临床特征,为婴幼儿急性腹泻的流行病学研究和临床治疗提供参考。方法选取2010至2014年苏浙闽地区多家医院门诊就诊的急性腹泻患儿1 452例,采用ELISA法检测A组轮状病毒(HRV),多重PCR法检测诺如病毒(NoV)、肠道腺病毒(EAdV)和星状病毒(HAstV),并用RT-PCR法对A组轮状病毒阳性的样本进行G和P分型。结果 1 452例患儿中由常见病毒引起的急性腹泻患儿有707例,占总调查患者的48.69%。急性腹泻患儿常见病毒感染中单一病毒感染占90.10%(637/707),其感染病毒的构成以轮状病毒最多(47.52%,336/707),其次为诺如病毒(36.07%,255/707)以及星状病毒(2.40%,17/707)和肠道腺病毒(4.10%,29/707)。病毒感染腹泻患儿中多重感染者共70例,占9.90%(70/707),以A组轮状病毒合并诺如病毒感染最为常见,占多重感染者的68.57%(48/70)。226份A组轮状病毒阳性标本中,G分型以G1为优势分型,占33.19%;P分型中则以P [11]比例最高,占38.50%;G、P型组合以P [11]G11为主(38.50%)。结论 A组轮状病毒是引发婴幼儿急性腹泻的主要病毒,诺如病毒次之,且婴幼儿病毒性腹泻患者可合并多种病毒感染。不同地区腹泻患儿病毒感染阳性率有差异,轮状病毒基因型表现则具有多样性。     

沙市婴幼儿腹泻中检出3株C群轮状病毒

1987～1988年在沙市165份婴幼儿急性腹泻标本中,用PAGE法检出轮状病毒37株(22.4％),其中3株为少见的轮状病毒,此种病毒经电镜观察,具有典型轮状病毒的形态结构,ELISA证实该病毒不具有A群和B群轮状病毒的群特异性抗原。RNA电泳分析表明,其基因组由11个双链RNA片段组成,电泳图型特殊,呈4:3:2:2的排列模式。上述试验表明,该病毒为世界上罕见的C群轮状病毒。免疫电镜证实,该病毒能被病人恢复期血清所凝集,提示该病毒是腹泻病儿的致病因子。     

双歧杆菌与利巴韦林治疗婴幼儿轮状病毒肠炎的疗效观察

秋冬季小儿腹泻的病原多是轮状病毒感染 ,临床多见于 6～ 2 4个月婴幼儿。我院 1 998年 1 1月至1 999年 1月腹泻病流行期间 ,用口服双歧杆菌 (丽珠肠乐 )与利巴韦林 (病毒唑 )泡滕剂联用治疗小儿轮状病毒肠炎 86例 ,疗效满意。1　对象与方法1 .1　病例选择　均系腹泻病流行期间急性腹泻患儿 ,病程在 3d以内 ,年龄 8～ 2 4个月 ,粪便均为稀水样或蛋花样 ,粪常规以消化不良、脂肪球为主 ,未检出红细胞、白细胞 ,大便培养阴性。采用金标免疫斑点法检测粪便 RV- Ag均阳性 ;根据 93年《中国腹泻病诊断治疗方案》诊断分型 ,均为轻至中型。随机分…     

腹泻患儿粪便A群轮状病毒抗原检测的结果分析

目的：了解本地区腹泻婴幼儿的A群轮状病毒感染情况及其流行特点。方法：采用胶体金法对我院2009年10月-2010年9月2104例有腹泻和肠炎特征的婴幼儿粪便进行A群轮状病毒抗原检测。结果：在2104例受检者中,A群轮状病毒感染的总阳性率是24．71％,其中男性感染率24．17％,女性为25．40％。不同年龄组间以1—2岁婴幼儿感染率最高,为32．13％,0-1岁为20．72％,2-5岁为12．03％。感染的季节特征是秋末冬初季（10—12月）阳性率最高,为42．82％,春末夏初季（4．6月）最低,为8．81％。结论：由A群轮状病毒感染引发的急性腹泻主要发生在1-2岁的婴幼儿,各个季节均有发生,以秋末冬初季为高发。     

猪和牛轮状病毒的分离与鉴定

应用MA-104细胞、胰蛋白酶处理培养物和旋转培养技术,成功地从患腹泻仔猪和犊牛的粪便样品中,分离到6株猪轮状病毒和4株牛轮状病毒。经形态学、血清学和病毒RNA电泳分析等鉴定,确为典型轮状病毒。经MA-104细胞连续传代,这10株轮状病毒均成为细胞适应毒株,并在该细胞上产生明显的细胞病变。     

Evidence for natural reassortants of human rotaviruses belonging to different genogroups.

Of 335 rotavirus isolates associated with diarrheal disease in Bangladesh that were culture adapted and subsequently characterized for electropherotype, subgroup, and serotype, 9 had properties that suggested they may be natural reassortants between human rotaviruses belonging to different "genogroups." Two of these were examined in greater detail by RNA-RNA hybridization with prototype strains representative of each of the three proposed human rotavirus genogroups. One subgroup II isolate, 248, with a "long" electrophoretic pattern was neutralized by hyperimmune antisera to both serotype 2 and 4 strains. Consistent with these results, seven RNA segments of this isolate formed hybrids with human strains belonging to the Wa genogroup and four segments hybridized with strains belonging to the DS-1 genogroup. The second isolate examined, 456, belonged to subgroup II and had a long electrophoretic pattern but was found to be a serotype 2 strain. This isolate also appeared to be an intergenogroup reassortant because three of its segments formed hybrids with strains belonging to the Wa genogroup and eight hybridized with viruses of the DS-1 genogroup. On the basis of the relative migration rates of these RNA-RNA hybrids during gel electrophoresis, a suggested origin for each gene segment was proposed which was consistent with the results expected from electrophoretic, subgroup, and serotypic analyses.     

Diagnosis of rotavirus infection with cloned cDNA copies of viral genome segments.

The diagnostic potential of cloned cDNA copies of human rotavirus (strain WA) genome segments for the detection of rotavirus in clinical specimens has been determined. A hybridization assay in which a mixture of 32P-labeled cDNAs representing the 11 rotavirus segments was used as a probe compared favorably with three frequently used diagnostic tests for rotavirus in terms of both specificity and sensitivity. Significantly, clinical isolates could be readily distinguished when cloned cDNA copies of individual genome segments were used independently as a probe. In assays in which genome RNA from rotaviruses of known subgroups and serotypes were tested, cloned probes that encode nonstructural viral proteins hybridized efficiently to genome RNAs of all strains, whereas cloned probes corresponding to genome segments 6 and 9 exhibited the potential for differentiating strains of different subgroups and serotypes. Cloned cDNA copies of rotavirus genome segments therefore offer considerable potential for improved general diagnosis of rotavirus in clinical specimens, as well as for epidemiological studies in which virus isolates can be distinguished on the basis of nucleotide sequence homology of individual genome segments.     

人轮状病毒的细胞培养分离

用CV-1细胞从急性腹泻病儿7份粪便标本中直接分离出4株人轮状病毒(HRV),并适应传代14代。感染细胞出现特征性细胞病变,经电镜、ELISA、免疫荧光染色及病毒RNA电泳等试验证实HRV毒株在CV-1细胞中的繁殖及抗原特异性。病毒滴度为10~(6.0)TCID_(60)。分离的4株HRV毒株均为RNA电泳型长型,经CV-1细胞传7～14代后,RNA图型与原粪样相比未见变异。     

Frequent reassortments may explain the genetic heterogeneity of rotaviruses: analysis of Finnish rotavirus strains

The predominant rotavirus electropherotypes (e-types) during 17 epidemic seasons (1980 through 1997) in Finland were established, and representative virus isolates were studied by nucleotide sequencing and phylogenetic analysis. The virus isolates were either P[8]G1 or P[8]G4 types. The G1 and G4 strains formed one G1 lineage (VP7-G1-1) and one G4 lineage, respectively. Otherwise, they belonged to two P[8] lineages (VP4-P[8]-1 and -2) unrelated to their G types. Phylogenetic analysis of partial sequences of all 11 RNA segments obtained from the strains also revealed genetic diversity among gene segments other than those defining P and G types. With the exception of segments 1, 3, and 10, the sequences of the other segments could be assigned to 2 to 4 different genetic clusters. The results of this study suggest that, in addition to the RNA segments encoding VP4 and VP7, the other RNA segments may segregate independently as well. In total, the 9 predominant e-types represented 7 different RNA segment combinations when the phylogenetic clusters of their 11 genes were determined. The extensive genetic diversity and number of e-types among rotaviruses are best explained by frequent genetic reassortment.     

Sequence diversity of human rotavirus strains investigated by northern blot hybridization analysis.

Rotavirus genomic RNAs, derived from a series of human isolates that exhibit variability in the pattern of migration of the double-stranded RNA on polyacrylamide gels, were transferred to diazobenzyloxymethyl paper, and their sequence diversity was investigated. Hybridization of cDNA probes prepared from the 11 segments of rotavirus RNA indicated that considerable sequence diversity exists among these viruses. Under conditions of both low and high stringency, hybridization analysis of virus collected between 1975 and 1980 suggested that the variation among rotavirus strains may have occurred by a process involving both "drift" and "shift" in the sequence of the rotavirus genomic segments.     

Distribution of human rotaviruses, especially G9 strains, in Japan from 1996 to 2000

A 4-year (1996-2000) survey of rotavirus infection involving 2,218 diarrheal fecal specimens of children collected from five regions of Japan was conducted. A total of 642 (28.9%) specimens were found to be rotavirus positive. A changed prevalence pattern of rotavirus G serotype was found with an increase of G9 and G2 and a decrease of G1, although G1 remained the prevailing serotype. Serotype G9 was unexpectedly determined to be the prevailing serotype in Sapporo (62.5%) and Tokyo (52.9%) in 1998-1999, and in Saga (78.4%) in 1999-2000. G9 strains isolated from 1998-1999 belonged to the P[8]-NSP4-Wa-group with long RNA pattern, while, G9 strains isolated from 1999-2000 belonged to three groups, the P[8]-NSP4-Wa-group with long RNA pattern, the P[4]-NSP4-KUN-group with short RNA pattern and a mixed-type group (P[4]/P[8]-NSP4-KUN/Wa-group with long RNA pattern). Both sequence and immunological analysis of VP7 revealed that the G9 strains from 1999-2000 were much more closely related to the G9 strains isolated worldwide in the 1990s, including G9 strains found in Thailand in 1997. However, the G9 strains from 1998-1999 were distinct from these and more closely related to the G9 prototype strains F45, AU32 and WI61 discovered in Japan and the US in the 1980s. Thus the G9 strains isolated in 1998-1999 had progenitors common to the G9 prototype strains, while the strains isolated in 1999-2000 did not directly evolve from them but were related to global G9 strains that have emerged in recent years. These data supported our previous report that G9 rotavirus might exist as two or more subtypes with diverse RNA patterns, P-genotype and NSP4 genogroup combinations (Y.M. Zhou et al., J. Med. Virol. 65: 619-628, 2001) and suggested that G9 rotavirus prevalent in Japan during two successive years belonged to different subtypes. The nucleotide sequences presented in this paper were submitted to DDBJ, EMBL and GenBank nucleotide sequence databases. The accession numbers are: 00-Ad2863VP7 (AB091746), 00-OS2986VP7 (AB091747), 00-SG2509VP7 (AB091748), 00-SG2518VP7 (AB091749), 00-SG2541 (AB091750), 00-SG2864 (AB091751), 00-SP2737VP7 (AB091752), 99-SP1542VP7 (AB091753), 99-SP1904VP7 (AB091754), 99-TK2082VP7 (AB091755) and 99-TK2091VP7 (AB091756).     

Detection of Rotavirus from Hospitalized Diarrheic Children in Uttar Pradesh, India

In the present study 220 stool samples collected from diarrheic children admitted to different hospitals and nursing homes of Uttar Pradesh and Uttarakhand were screened for rotavirus. Of 220 diarrheic samples screened 46 samples were found to be positive for rotavirus by RNA PAGE. All the isolates exhibited 4-2-3-2 migration pattern suggesting group A rotavirus. Both long and short electropherotypes were prevalent in these regions. Six different electropherotypes were detected in this study period. Male diarrheic children were found to be more susceptible to rotavirus infection (22.96 %) than that of the female ones (17.64 %). Viral RNA isolated from stool samples again subjected to VP4 gene amplification by RT-PCR using con2 and con3 primer which resulted 876 bp product suggesting group A rotavirus. Besides virus isolation was successfully done using MA104 cell line.     

The nature of virus strains isolated from patients with poliomyelitis in the Byelorussian SSR 1967-1985

Oligonucleotide mapping of the RNA genome was applied for analysis of poliovirus clinical isolates in Byelorussia in 1967-1985. All the maps have been clearly shown to resemble the reference maps of the corresponding vaccine strains. Examination of the oligonucleotide maps of poliovirus isolates provides a measure of distribution of some of the mutations over the viral genome.