目的 分析钩端螺旋体(简称钩体)疫苗部分菌种的分子特性及毒力。方法 应用多位点可变数量串联重复序列分析(multi locus variable number tandem repeat analysis,MLVA)方法,结合脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE...目的 分析钩端螺旋体(简称钩体)疫苗部分菌种的分子特性及毒力。方法 应用多位点可变数量串联重复序列分析(multi locus variable number tandem repeat analysis,MLVA)方法,结合脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)技术,分析我国4株钩体疫苗弱毒株的分子特性;通过豚鼠动物模型攻击实验分析4株弱毒株的毒力。结果 4株钩体疫苗弱毒株经MLVA分析呈现4种不同的型;PFGE分析显示,经NotⅠ酶切,染色体呈现8~13个长度不等的片段;所有经弱毒株攻击动物的主要脏器均出现典型的钩体感染病变。结论 明晰了4株钩体疫苗弱毒株的分子特性及其对豚鼠的毒力,为完善该疫苗菌种的质量控制积累了资料。展开更多
Objective To prepare the 4 candidate vaccine strains of H5N1 avian influenza virus isolated in China Methods Recombinant viruses were rescued using reverse genetics. Neuraminidase （NA） and hemagglutinin （HA） segme...Objective To prepare the 4 candidate vaccine strains of H5N1 avian influenza virus isolated in China Methods Recombinant viruses were rescued using reverse genetics. Neuraminidase （NA） and hemagglutinin （HA） segments of the A/Xinjiang/1/2006, A/Guangxi/1/2009, A/Hubei/1/2010, and A/Guangdong/1/2011 viruses were amplified by RT-PCR. Multibasic amino acid cleavage site of HA was removed and ligated into the pCIpoll vector for virus rescue. The recombinant viruses were evaluated by trypsin dependent assays. Their embnjonate survival and antigenicity were compared with those of the respective wild-type viruses. Results The 4 recombinant viruses showed similar antigenicity compared with wild-type viruses, chicken embryo survival and trypsin-dependent characteristics. Conclusion The 4 recombinant viruses rescued using reverse genetics meet the criteria for classification of low pathogenic avian influenza strains, thus supporting the use of them for the development of seeds and production of pre-pandemic vaccines.展开更多
文摘目的 分析钩端螺旋体(简称钩体)疫苗部分菌种的分子特性及毒力。方法 应用多位点可变数量串联重复序列分析(multi locus variable number tandem repeat analysis,MLVA)方法,结合脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)技术,分析我国4株钩体疫苗弱毒株的分子特性;通过豚鼠动物模型攻击实验分析4株弱毒株的毒力。结果 4株钩体疫苗弱毒株经MLVA分析呈现4种不同的型;PFGE分析显示,经NotⅠ酶切,染色体呈现8~13个长度不等的片段;所有经弱毒株攻击动物的主要脏器均出现典型的钩体感染病变。结论 明晰了4株钩体疫苗弱毒株的分子特性及其对豚鼠的毒力,为完善该疫苗菌种的质量控制积累了资料。
基金supported by the National High Technology Research and Development Program of China(863 Program)SQ2009AA02XK1487370
文摘Objective To prepare the 4 candidate vaccine strains of H5N1 avian influenza virus isolated in China Methods Recombinant viruses were rescued using reverse genetics. Neuraminidase （NA） and hemagglutinin （HA） segments of the A/Xinjiang/1/2006, A/Guangxi/1/2009, A/Hubei/1/2010, and A/Guangdong/1/2011 viruses were amplified by RT-PCR. Multibasic amino acid cleavage site of HA was removed and ligated into the pCIpoll vector for virus rescue. The recombinant viruses were evaluated by trypsin dependent assays. Their embnjonate survival and antigenicity were compared with those of the respective wild-type viruses. Results The 4 recombinant viruses showed similar antigenicity compared with wild-type viruses, chicken embryo survival and trypsin-dependent characteristics. Conclusion The 4 recombinant viruses rescued using reverse genetics meet the criteria for classification of low pathogenic avian influenza strains, thus supporting the use of them for the development of seeds and production of pre-pandemic vaccines.
