摘要
目的对A组轮状病毒武汉流行毒株进行培养增殖并确定其基因型和血清型。材料经RT-PCR确定基因型和ELISA确定VP6亚组血清型后初步判定的57株不常见型、混合型及未确定型别的A组轮状病毒流行毒株。方法MA104细胞分离培养病毒;聚丙烯酰胺凝胶电泳确定电泳型;逆转录巢式聚合酶链反应确定G、P、VP6和NSP4基因型;ELISA确定VP6亚组血清型。结果分离率77.2%(44/57)。分离了一株具备双重亚组血清型特征的人轮状病毒R479。分离后基因混合型、非常见基因型别或非常见G、P组合、不能确定的基因型别及VP6亚组血清混合型的样本数量与分离前相比显著减少。结论轮状病毒流行毒株的分离培养有助于提高其基因分型的敏感性,减少基因分型和血清分型的非特异性反应。
The 44 field strains of human rotavirus were isolated through cultivation of virus on MA-104 cells from 57 fecal specimens of patients with acute diarrhea from 2000 to 2005. Rotaviruses isolated from these specimens were preliminarily analyzed for their G, P,VP6, NSP4 genotypes by RT-PCR and subgrouped by ELISA with specific monoclonal antibodies against subgroup （SG）1 and II. The results showed that the isolation rate was found to be 77.2 % （44/57）, including one of G9 strain, two of [8] ＋ [4] strains, one of G4 SG Ⅰ＋ Ⅱstrain designated as strain R479, one strain with double 6th bands in PAGE as well. Presence of all the isolated rotaviruses from cultures was confirmed by the detection of RNA pattern by polyacrylamide gel electrophoresis . Before and after tissue cultivation of rotaviruses, there were 7 and 2 P-untypable strains, 8 and 3 strains of mixed G types, 22 and 4 strains of mixed P types, 17 and 1 strains of SG Ⅰ＋Ⅱ type, respectively. From these observation, it is apparent that tissue cultures of rotaviruses is helpful to improve the sensitivity of genotyping and reducing the presence of non-specific reactions of serotyping in molecular epidemiological studies of rotavirus.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2008年第10期919-922,共4页
Chinese Journal of Zoonoses
基金
日本文部省2006-2009年度科研辅助金(No.18406018)
厚生省国际健康合作研究基金(No.18-5)联合资助
关键词
轮状病毒
A组
野毒株
培养
鉴定
rotavirus
group A
field strains
cultivation
identification
