| 黄永坤,戚勤,侯宗柳,李海林,文革生,庞伟,周丽芳.昆明地区22株人轮状病毒VP7及NSP4基因分析[J].Chinese journal of Epidemiology,2005,26(12):980-983 |
| 昆明地区22株人轮状病毒VP7及NSP4基因分析 |
| Analysis on molecular characteristic of VP7and NSP4 |
| Received:February 05, 2005 |
| DOI： |
| KeyWord: 轮状病毒 VP7 NSP4 基因型 |
| English Key Word: Rotavirus VP7 NSP4 Serotype |
| FundProject:云南省教育厅重点科研基金(03Z442C) |
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| Abstract: |
| 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、G4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、NUN、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RV VP7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。 |
| English Abstract: |
| Objective To explore the molecular characteristics and molecular variation of human rotavirus(HRV) strains and to understand the relationship between clinical characteristics and epidemiology of different HRV-VP7 and NSP4.Methods Double-strand RNA of rotavirus extracted from stool samples was used as the template for reverse transcription of gene VP7, which was followed by nested PCR for VP7 typing.NSP4 genes from 22 epidemic strains of human rotavirus isolated in Kunming in 2002 and 2003 were amplified with RT-PCR.Then cDNAs were sequeneed and compared with 4 human rotavirus NSP4 (Wa,KUN,AU-1,Hochi)) and 3 animal rotavirus NSP4 (EW, OSU, SA11) available in the GenBank while the epidemic strains of human rotavirus isolated in different areas of China were compared, using the Clustal-mp,DNAssist,MEGA2 software.The G serotype of VP7 was analysed by PCR.Results Serotype G1 was prevalent in 2002 while serotype G3 was the prevalent in Kumming in 2003.The NSP4 genes from 22 epidemic strains of human rotavirus isolated in Kunming in 2002 and 2003 belonged to Wa with highly conservative amino acid.Samples isolated in the same years but not in the same area shared higher homology.Symptoms associated with heavy diarrhea did not seem to be associated with NSP4 molecular variation(P0.05).Conclusion Obvious variations of VP7 typing were seen in the same season, as well as in different areas and years.Due to the stable nature of NSP4, it seem to be a better candidate for vaccine production, than VP7. |
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