This paper reports the confirmed diagnosis by nested RT-PCR of PPR cases in Tibet, China in 2007, and results of phylogenetic analysis. Results showed that the 11 tested samples were PPRV positive by nested RT-PCR, of...This paper reports the confirmed diagnosis by nested RT-PCR of PPR cases in Tibet, China in 2007, and results of phylogenetic analysis. Results showed that the 11 tested samples were PPRV positive by nested RT-PCR, of which 2 samples were genetically close to the X7443 strain (Nigeria 75/1) of lineage I, and 3 samples close to the strain AY560591 (Sungri96) of linage IV with 96.6%、97.3%、97.6% and 98% nucleotide sequence homogeneity respectively, based on partial sequencing of the F gene from 5 samples and complete sequencing of the N/M/F/H genes from one sample. This study suggested that there are at least 2 origins of PPRV in China.展开更多
Peste des petits ruminants(PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding...Peste des petits ruminants(PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding of PPR virus(PPRV)pathobiology and molecular biology is critical for effective control and eradication of the disease. To achieve these goals,establishment of stable reverse genetics systems for PPRV would play a key role. Unfortunately, this powerful technology remains less accessible and poorly documented for PPRV. In this review, we discussed the current status of PPRV reverse genetics as well as the recent innovations and advances in the reverse genetics of other non-segmented negative-sense RNA viruses that could be applicable to PPRV. These strategies may contribute to the improvement of existing techniques and/or the development of new reverse genetics systems for PPRV.展开更多
为了建立一种快速、特异、敏感的小反刍兽疫病毒(peste des petits ruminants virus,PPRV)血清学检测方法,本研究将PPRV血凝蛋白(H)和核蛋白(N)克隆至pET28a(+)载体进行诱导表达,并将2种蛋白Ni柱纯化后包被ELISA反应板,建立起检测PPRV...为了建立一种快速、特异、敏感的小反刍兽疫病毒(peste des petits ruminants virus,PPRV)血清学检测方法,本研究将PPRV血凝蛋白(H)和核蛋白(N)克隆至pET28a(+)载体进行诱导表达,并将2种蛋白Ni柱纯化后包被ELISA反应板,建立起检测PPRV特异性抗体的间接ELISA法。结果表明在E.coli BL21(DE3)中表达出2种PPRV蛋白,将2种重组蛋白纯化后按照1:1比例混合,制备PPRV鸡尾酒抗原,成功建立了特异性强、敏感性高的PPRV间接ELISA法;最后用建立的方法对新疆伊犁、塔城、阿勒泰和喀什边境地区采集的325份山羊、789份绵羊和132份牛血清进行了抗体检测,检测结果显示均为阴性,提示在我国新疆边境地区反刍动物尚未发生PPRV感染,但需要及时进行免疫接种以建立预防PPRV从国外传入的免疫带。本研究建立的PPRV间接ELISA法为该病毒的检测提供基础,也为该病的科学防控提供参考。展开更多
为探讨小反刍兽疫病毒(peste des petits ruminants virus,PPRV)贵州流行株N基因分子特征和分群,试验设计了1对特异性引物,应用RT-PCR技术对小反刍兽疫(peste des petits ruminants,PPR)临床样本进行N基因扩增,克隆至pMD19-T载体,对阳...为探讨小反刍兽疫病毒(peste des petits ruminants virus,PPRV)贵州流行株N基因分子特征和分群,试验设计了1对特异性引物,应用RT-PCR技术对小反刍兽疫(peste des petits ruminants,PPR)临床样本进行N基因扩增,克隆至pMD19-T载体,对阳性重组质粒进行测序,应用DANStar软件对测序序列和参考序列进行核苷酸同源性、氨基酸同源性、变异位点及系统进化树分析。结果显示:PPRV贵州流行株N基因扩增长度为1 578bp,其相互间核苷酸、氨基酸同源性分别为99.6%~100.0%及99.2%~100.0%,与国内参考株N基因的核苷酸序列(97.7%~99.9%)及氨基酸序列(98.3%~100.0%)同源性较国外参考株(88.5%~97.7%和92.2%~98.5%)高;PPRV贵州流行株N基因编码的氨基酸同疫苗株Nigeria 75-1相比存在26个位点突变,但没有氨基酸的缺失或增加;基于N基因系统进化分析显示,PPRV贵州流行株同国内参考株处于同一个进化分支,但与国外参考株处于不同进化分支;其属于病毒进化的Ⅳ基因群,与国内参考株处于同一系统分群,但与疫苗株Nigeria 75-1(Ⅰ基因群)处于不同基因群。展开更多
基金This work was supported by project from MOA [2006-G57(3)B-Z1] Project from Yunnan Province (2008LA019)
文摘This paper reports the confirmed diagnosis by nested RT-PCR of PPR cases in Tibet, China in 2007, and results of phylogenetic analysis. Results showed that the 11 tested samples were PPRV positive by nested RT-PCR, of which 2 samples were genetically close to the X7443 strain (Nigeria 75/1) of lineage I, and 3 samples close to the strain AY560591 (Sungri96) of linage IV with 96.6%、97.3%、97.6% and 98% nucleotide sequence homogeneity respectively, based on partial sequencing of the F gene from 5 samples and complete sequencing of the N/M/F/H genes from one sample. This study suggested that there are at least 2 origins of PPRV in China.
基金supported by the National Key Research and Development Program of China (2016YFD0500108 and 2016YFE0204100)the International Cooperation Project of CAAS Innovation Program (CAAS-GJHZ201700X)
文摘Peste des petits ruminants(PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding of PPR virus(PPRV)pathobiology and molecular biology is critical for effective control and eradication of the disease. To achieve these goals,establishment of stable reverse genetics systems for PPRV would play a key role. Unfortunately, this powerful technology remains less accessible and poorly documented for PPRV. In this review, we discussed the current status of PPRV reverse genetics as well as the recent innovations and advances in the reverse genetics of other non-segmented negative-sense RNA viruses that could be applicable to PPRV. These strategies may contribute to the improvement of existing techniques and/or the development of new reverse genetics systems for PPRV.
