Zeng Mei, Zhu Qi-rong, Zhang You, Li Guo-hua, Chen Dong-mei, Ding Ya-xin, Qian Yuan
Children's Hospital, Fudan University, Shanghai, 200032 China.
Zhonghua Er Ke Za Zhi. 2004 Jan;42(1):10-5.
To investigate molecular epidemiologic features of rotaviruses circulating in Shanghai, China.
Stool samples were collected from 1230 hospitalized children with community-acquired and nosocomially acquired diarrhea in Children's Hospital Affiliated to Fudan University between November 1, 1999 and December 31, 2001. Polyacrylamide gel electrophoresis (PAGE) was used to detect rotavirus genomic RNA and identify electropherotypes of group A rotavirus RNAs. Reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify full length VP7 gene and dot blot hybridization was performed to identify rotavirus G serotypes using digoxigenin-labelled variable regions from VP7 genes as probes. These probes were amplified by PCR from recombinant plasmids containing full length G1, G2, G3 and G4 VP7 genes from rotavirus field strains detected in Beijing and digoxigenin labelled dUTP was integrated into the PCR products. The Kruskal-Wallis analysis of variance was employed to analyze whether there were significant differences in variables.
Out of 1230 samples investigated, 493 (40.1%) were group A rotavirus gene positive by PAGE, among which 397 (80.5%) showed long electropherotypes, 55 (11.2%) showed short electropherotypes, 18 (3.7%) showed mixed electropherotypes which suggested that the children were co-infected by rotaviruses with different electropherotypes, 23 (4.7%) were non-typable because of degradation of some of the genomic RNA fragments. No group B or group C rotavirus was found. RT-PCRs were performed for 328 fecal specimens containing sufficient rotavirus RNAs and VP7 gene products were obtained from 254 (77.4%) samples. Dot blot hybridization showed serotype G1 accounted for 55.5% (141) of these samples, serotype G3 accounted for 27.6% (70), serotype G2 accounted for 9.4% (24), co-infection by 2 rotaviruses with different G types accounted for 6.3% (16), only 1 G4 was detected and 2 were non-typable. The genomic RNA patterns of all G2 strains were short and those of G1, G3 and G4 strains were long. There were no statistically significant differences for age distribution and clinical manifestations among those infants and children infected by rotaviruses with different G serotypes.
Group A rotavirus is the major pathogen for diarrhea in infants and children in Shanghai during the period of Nov. 1999 to Dec. 2001. Rotaviruses with long electropherotype were dominant during these years. Serotypes G1 to G3 constituted 98.8% of all 254 strains tested, and G1 was the most common serotype followed by G3 and G2, whereas serotype G4 was seldom found. Some of the children were co-infected by rotaviruses with different G serotypes. Clinical manifestations were not related to the infecting rotavirus with different G serotypes.
研究中国上海地区流行的轮状病毒的分子流行病学特征。
收集1999年11月1日至2001年12月31日期间复旦大学附属儿科医院1230例社区获得性和医院获得性腹泻住院患儿的粪便样本。采用聚丙烯酰胺凝胶电泳（PAGE）检测轮状病毒基因组RNA并鉴定A组轮状病毒RNA的电泳型。进行逆转录聚合酶链反应（RT-PCR）扩增全长VP7基因，并用地高辛标记的VP7基因可变区作为探针进行斑点杂交以鉴定轮状病毒G血清型。这些探针通过PCR从含有在北京检测到的轮状病毒野毒株全长G1、G2、G3和G4 VP7基因的重组质粒中扩增得到，地高辛标记的dUTP整合到PCR产物中。采用Kruskal-Wallis方差分析来分析变量之间是否存在显著差异。
在1230份调查样本中，493份（40.1%）经PAGE检测为A组轮状病毒基因阳性，其中397份（80.5%）显示长电泳型，55份（11.2%）显示短电泳型，18份（3.7%）显示混合电泳型，提示患儿被不同电泳型的轮状病毒共同感染，23份（4.7%）因部分基因组RNA片段降解而无法分型。未发现B组或C组轮状病毒。对328份含有足够轮状病毒RNA的粪便标本进行RT-PCR，254份（77.4%）样本获得了VP7基因产物。斑点杂交显示G1血清型占这些样本的55.5%（141份），G3血清型占27.6%（70份），G2血清型占9.4%（24份），两种不同G型轮状病毒共同感染占6.3%（16份），仅检测到1份G4型，2份无法分型。所有G2毒株的基因组RNA模式均为短型，G1、G3和G4毒株的基因组RNA模式为长型。不同G血清型轮状病毒感染的婴幼儿在年龄分布和临床表现方面无统计学显著差异。
1999年11月至2001年12月期间，A组轮状病毒是上海地区婴幼儿腹泻的主要病原体。这些年中长电泳型轮状病毒占主导。G1至G3血清型占所有检测的254株毒株的98.8%，其中G1是最常见的血清型，其次是G3和G2，而G4血清型很少见。部分儿童被不同G血清型的轮状病毒共同感染。临床表现与感染不同G血清型的轮状病毒无关。
