中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2010年
27期
1924-1928
,共5页
谌资%郑煜煌%Yipu Lin%Patrick J Collins%Alan J Hay
諶資%鄭煜煌%Yipu Lin%Patrick J Collins%Alan J Hay
심자%정욱황%Yipu Lin%Patrick J Collins%Alan J Hay
正粘病毒科%神经氨酸酶%突变%酶抑制剂%禽流感病毒H5N1
正粘病毒科%神經氨痠酶%突變%酶抑製劑%禽流感病毒H5N1
정점병독과%신경안산매%돌변%매억제제%금류감병독H5N1
Orthomyxoviridae%Neuraminidase%Mutation%Enzyme inhibitors: Avian influenza virus H5N1
目的 研究感染人的禽流感病毒H5N1神经氨酸酶突变I117V,I314V,I117V+I314V对神经氨酸酶抑制剂(NAIs)敏感性和神经氨酸酶(NA)活性的影响.方法 定点突变技术将变异位点引入禽流感病毒株A/Vietnam/1203/04(H5N1)(VN1203)的NA基因中,以A/WSN/33(H1N1)为背景,通过反向遗传学方法构建重组流感病毒,鸡胚传代增殖病毒,荧光定量评估病毒对NAIs的敏感性和NA活性(IC50值和Km,Ki值).结果 与野生株VN1203相比,I117V使NA对奥司他韦敏感性降低(IC50值升高了17倍,Ki值升高了20倍),对扎那米韦敏感性无明显影响(IC50值升高了2倍,Ki值升高了3倍),NA活性降低(Km值升高了23倍).I314V对NAIs敏感性和NA活性均无明显影响.结论 初步结果认为NA突变I117V和I314V对感染人的禽流感病毒H5N1并不产生NAIs耐药,临床上仍然可以用奥司他韦或扎那米韦抗病毒治疗.
目的 研究感染人的禽流感病毒H5N1神經氨痠酶突變I117V,I314V,I117V+I314V對神經氨痠酶抑製劑(NAIs)敏感性和神經氨痠酶(NA)活性的影響.方法 定點突變技術將變異位點引入禽流感病毒株A/Vietnam/1203/04(H5N1)(VN1203)的NA基因中,以A/WSN/33(H1N1)為揹景,通過反嚮遺傳學方法構建重組流感病毒,鷄胚傳代增殖病毒,熒光定量評估病毒對NAIs的敏感性和NA活性(IC50值和Km,Ki值).結果 與野生株VN1203相比,I117V使NA對奧司他韋敏感性降低(IC50值升高瞭17倍,Ki值升高瞭20倍),對扎那米韋敏感性無明顯影響(IC50值升高瞭2倍,Ki值升高瞭3倍),NA活性降低(Km值升高瞭23倍).I314V對NAIs敏感性和NA活性均無明顯影響.結論 初步結果認為NA突變I117V和I314V對感染人的禽流感病毒H5N1併不產生NAIs耐藥,臨床上仍然可以用奧司他韋或扎那米韋抗病毒治療.
목적 연구감염인적금류감병독H5N1신경안산매돌변I117V,I314V,I117V+I314V대신경안산매억제제(NAIs)민감성화신경안산매(NA)활성적영향.방법 정점돌변기술장변이위점인입금류감병독주A/Vietnam/1203/04(H5N1)(VN1203)적NA기인중,이A/WSN/33(H1N1)위배경,통과반향유전학방법구건중조류감병독,계배전대증식병독,형광정량평고병독대NAIs적민감성화NA활성(IC50치화Km,Ki치).결과 여야생주VN1203상비,I117V사NA대오사타위민감성강저(IC50치승고료17배,Ki치승고료20배),대찰나미위민감성무명현영향(IC50치승고료2배,Ki치승고료3배),NA활성강저(Km치승고료23배).I314V대NAIs민감성화NA활성균무명현영향.결론 초보결과인위NA돌변I117V화I314V대감염인적금류감병독H5N1병불산생NAIs내약,림상상잉연가이용오사타위혹찰나미위항병독치료.
Objective To study the impact of avian influenza virus H5N1 neuraminidase mutations
I117V,I314V and I117V+I314V on the sensibility of neuraminidase inhibitors(NAIs) and the activity of neuraminidase (NA).Methods The mutations were intreduced into NA genes of virus strain A/Vietnam/1203/04 (H5N1)by site-directed mutagenesis.With the A/WSN/33 (H1N1)background,recombinant influenza viruses containing NA mutations were rescued by reverse genetics.After viral propagation in chicken embryos,fluorimetric assays were conducted to assess the sensibility to NAIs and NA activity(IC50,Km&Ki).Results Compared to the wild-type viruS VNl203.the mutation I117V decreased the susceptibility to osehamivir(17-fold increment of IC50 value,20-fold increment of Ki value)and the NA activity(23-fold increment of Km value)while there Was little impact on zanamivir sensitivity(2-fold increment of IC50 value,3-fold increment of Ki value).The mutation 1314V had no marked influence on either the NA activity or the NAIs susceptibility.Conclusion It aDpears that the NA mutations of I117V and I314V Can not cause NAIs resistanee.Oseltamivir or zanamivir may still be prescribed for anti-viral treatment.
