天津地区住院腹泻儿童轮状病毒的检测及其毒株型别分析

目的 了解天津地区5岁以下住院腹泻患儿A组RV的感染情况及其型别特点.方法 收集天津市儿童医院2008年5月至2009年4月837份住院腹泻患儿的粪便标本,用胶体金免疫层析法快速检测A组RV抗原,将检测阳性的标本进行细胞培养,产生细胞病变(CPE)后,提取病毒RNA,RT-PCR扩增病毒VP7基因,并将PCR产物阳性标本测序,证实其VP7(G)血清型.同时收集其临床相关资料.结果 837份标本RV抗原阳性率为26.3%(220/837);90.5%(199/220)的RV腹泻发生在24月龄以下患儿;发病高峰时间主要集中在2008年10月至2009年4月;208份接种的RV抗原阳性标本中,病毒分离后用PCR鉴定阳性95份,对其中35份电泳较好的PCR产物阳性标本作序列分析,除1份为G9型、2份为G3型外,其余均为RV G1型.结论 RV是天津地区婴幼儿腹泻的重要病原,流行的基因型以G1为主.

Detection and genotyping of rotavirus among children under 5 years old hospitalized with diarrhea in Tianjin

Objective To study the prevalence and genotypes of rotavirus (RV) among children,< 5 years old hospitalized with viral diarrhea in Tianjin. Methods Stool specimens were collected from hospitalized diarrhea children in Tianjin children's hospital between May 2008 and April 2009. Detection of rotavirus was employed by Colloidal Gold Device. The detected positives were inoculated to MA-104 cells. The total RNA of virus was extracted after CPE which was caused by rotavirus were observed, The VP7 serotypes were determined by using RT-PCR to amplify the VP7 gene and sequencing the RT-PCR products.The clinical data for each patient were also collected. Results Among 837 specimens, the RV antigen positive rate was 26. 3% (220/837). Among all the children with rotavirus diarrhea, 90. 5% (199/220)were < 2 years old. The prevalence of rotavirus diarrhea in children peaked during Oct. 2008 through Apr.2009. Of the 208 rotavirus positive specimens, 95 were successfully identified by RT-PCR Thirty-five positive strains of RV were sequenced, and the sequencing results showed that 32 positive strains were belonged to rotavirus G1 type, 2 positive strains were belonged to rotavirus G3 type and 1 positive strain were belonged to rotavirus C9 type. Conclusion RV was the dominant etiological agent for infantile diarrhea infection in Tianjin, and the predominant serotype was G1.