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1.
1981年,美国费城维斯特研究院Anilionis等首先报告了克隆狂犬病毒ERA株抗原基因,并获得无性繁殖成功,从而测定了抗原糖蛋白基因的序列,推导出它的氨基酸排列。1983年,美国基因技术公司Yelverton等又测定了狂犬病毒CVS株糖蛋白基因序列,进一步合成了克隆引物和探针十五核酸片段5′CATGAAGTTCCCCAT 3′。由该片 相似文献
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本文利用聚合酶链式反应对来自澳大利亚昆士兰州、维多利亚州和新南维尔士州的４株禽痘病毒田间分离株及二疫苗株中网状内皮组织增殖病毒的ＬＴＲ片断、ｅｎｖ及ｒｅｌ基因因进行了检测。所有４株痘病毒分离株均为ＬＴＲ片断、ｅｎｖ阳性，曾被ＰＣＲ证实为ＬＴＲ阳性的一疫苗株在本实验中也为ＬＴＲ、ｅｎｖ阳性，另一疫苗株为ＬＴＲ、ｅｎｖ阴性，所有上述禽痘病毒株均为ｒｅｌ阴性。 相似文献
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狂犬病毒、伪狂犬病毒、马瘟病毒、牛病毒性腹泻／粘膜病病毒和传染性牛鼻气管炎病毒的保存期试验 总被引：1，自引：1，他引：0
进行了狂犬病毒（巴黎株固定毒）、伪狂犬病毒（闽A株，苏联株）、马瘟病毒（Ⅰ，Ⅱ，Ⅲ，Ⅳ，Ⅴ，Ⅶ型）、牛病毒性腹泻／粘膜病病毒（Oregon C24V株，NADL株）和传染性牛鼻气管炎病毒（NU／67株）的保存期试验。试验结果证明，在－70℃以下温度保存，狂犬病毒巴黎株固定毒保存10年以上，伪狂犬病毒闽A株保存11年以上，伪狂犬病毒苏联株保存保存17年，马瘟病毒Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ和Ⅶ型病毒保存12年以上，牛病毒性腹泻／粘膜病病毒NADL株和Oregon C24V株保存16年以上，传染性鼻鼻气管炎病毒NU／67株保存14年以上均未见病毒滴度的显著下降。 相似文献
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H5亚型禽流感重组鸡痘病毒活载体疫苗 总被引：1，自引：0，他引：1
H5亚型禽流感重组鸡痘病毒活载体疫苗系以高度成熟安全的鸡痘病毒为载体研制的新型基因工程禽流感一鸡痘二联疫苗，该疫苗毒株的研制是采用重组DNA技术，将我国H5N1亚型禽流感病毒早期分离株的HA和NA基因同源重组到鸡痘病毒疫苗株的基因组中，构建重组鸡痘病毒(rFPV—HA—NA)，经体外、体内试验表明该重组鸡痘病毒具有良好的免疫原性。同时经细胞连续传代，证实了重组鸡痘病毒具有良好的遗传稳定性，从而进一步保证了该疫苗的免疫原性。 相似文献
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将禽流感病毒A／Goose／Guangdong／1／96（H5N1）株的NA基因及LacZ报告基因克隆到禽痘病毒载体pSY681中，构建重组转移载体pSY（NA＋LacZ），将其转染鸡胚成纤维细胞，经蓝白斑筛选，纯化表达NA基因的重组禽痘病毒rFPV-NA，用其免疫SPF鸡，检测该重组禽痘病毒的免疫原性。结果显示，该重组禽痘病毒能够在体外培养细胞内表达具有生物学活性的NA蛋白，表达产物的分子质量约为55ku；用该重组禽痘病毒免疫SPF鸡后，能够使免疫鸡获得对H5N1和H7N1两种亚型HPAIV攻击的部分保护。表明，重组禽痘病毒rFPV—NA具有良好的免疫原性。 相似文献
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将鸡传染性支气管炎病毒S1基因插入到鸡痘病毒转移载体pSY681中，获得重组转移载体pSY681。将pSY681-IBVS1转染已感染亲本鸡痘病毒S-FPV-017株的鸡胚成纤维细胞，使其在鸡胚成纤维细胞内与鸡痘病毒基因组发生同源重组，产生表达鸡IBVS1蛋白的重组鸡痘病毒rFPV-IBVS1。在含有X-gal的营养琼脂培养基上进行蓝斑筛选且进一步纯化14代。S1基因的PCR检测表明，获得的含传染性支气管炎病毒S1基因的重组鸡痘病毒能够稳定遗传，间接免疫荧光和Western blot等试验证实该重组病毒在CEF内真实地表达了分子量约为90Ku的具有免疫学活性的IBV S1糖蛋白。 相似文献
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本文利用聚合酶链式反应对来自澳大利亚昆士兰州,维多利亚州的新南维尔士州的４株禽痘病毒田间分离株及二疫苗株中网状内皮组织增殖病毒的ＬＴＲ片断,ｅｎｖ及ｒｅｌ基因因进行了检测。 相似文献
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本试验利用PK15细胞从江西省某猪场疑似猪痘的保育猪皮肤痘痂中分离到1株病毒,该分离毒株在PK15单层细胞上能连续盲传培养,不产生明显的细胞病变,通过PCR扩增猪痘病毒P35基因,扩增序列与猪痘病毒参考株(AF410153.1)P35基因的核苷酸同源性为97.9%,表明该分离病毒为猪痘病毒,并命名为SWPV-JX01。将SWPV-JX01株F5代细胞培养液经肌肉和皮下注射感染兔和小鼠,人工感染80 h后,通过PCR检测不同组织的猪痘病毒P35基因,从兔的肾脏、脾脏、皮肤、心肌、淋巴结均能扩增到P35基因,且扩增序列与SWPV-JX01株同源性为100%。试验结果表明,SWPV-JX01株能在兔体内增殖,但不能在昆明鼠体内增殖。 相似文献
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江西某猪场的部分保育猪皮肤出现数量不等的痘斑,为确定其病原,根据猪痘病毒(AF410153.1)设计1对引物,以该场发病猪的皮肤痘痂提取DNA,采用PCR的方法扩增出与预期大小相符的目的条带,测序结果表明该扩增片段为猪痘病毒核酸序列,与猪痘病毒参考毒株(AF410153.1)的同源性为98%。利用PK15细胞盲传从病猪皮肤痘痂中分离到一株猪痘病毒,该分离株在PK15细胞上不产生细胞病变(CPE)。采用间接ELISA方法对该场的24份猪血样进行猪痘病毒抗体检测,结果猪痘抗体阳性血清数12份(阳性率50%)。 相似文献
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Arathy DS Tripathy DN Sabarinath GP Bhaiyat MI Chikweto A Matthew V Sharma RN 《Avian diseases》2010,54(3):1081-1085
Two 1-mo-old local breed chickens, with gross lesions in the skin of the head region suspected to be fowl poxvirus infection, were submitted to the Diagnostic Laboratory of the School of Veterinary Medicine, Grenada, West Indies. Cutaneous lesions were collected from these birds for virus isolation, histopathologic diagnosis, and molecular analysis. Fowl poxvirus infection was confirmed by virus isolation in chicken embryo and by histopathology. Molecular characterization of the fowl poxvirus was conducted by PCR amplification of selected genomic fragments and by nucleotide sequencing. Integration of reticuloendotheliosis virus fragments into the fowl poxvirus genome was confirmed by PCR and DNA sequencing. This is the first report from the Caribbean region on the preliminary molecular characterization of a fowl poxvirus isolate. 相似文献
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Hess C Maegdefrau-Pollan B Bilic I Liebhart D Richter S Mitsch P Hess M 《Avian diseases》2011,55(4):714-718
The present report documents the occurrence of a poxvirus infection in commercial meat turkeys. The affected farm had six flocks, with a total of 11,680 birds at different ages; birds from two of these flocks were affected. The clinical picture was characterized by severe epithelial lesions and proliferations on the head and neck regions as reported for the cutaneous form of poxvirus infection. Except for these lesions, no adverse clinical signs or gross pathologic lesions were observed. Only a low number of birds was affected (n = 20) and no increase of mortality could be seen. Bacteriologic investigations from the lesions revealed multiresistant Staphylococcus aureus. Eosinophilic inclusions (Bollinger bodies) in histologic examinations in the cytoplasm of keratinocytes were noticeable. Typical pox virions were demonstrated by electron microscopy, and poxvirus was isolated on the chorioallantoic membrane of specific-pathogen-free chicken eggs. Further identification of the poxvirus species was carried out by PCR and sequencing, revealing an infection with the species fowlpox. Layers in vicinity of the turkey farm that also were affected by fowlpox were considered as potential source of infection. Although it is assumed that avian poxviruses are strongly species specific, the present case report reinforces the changing picture of poxvirus infections in turkeys. Furthermore, it supports the assumption of previous data that fowlpox virus has to be seen as recently emerging pathogen in turkeys. 相似文献
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An avian poxvirus from the beak scab of an American flamingo (Phoeniconais ruber rubber) was isolated by inoculation on the chorioallantoic membrane (CAM) of specific-pathogen-free (SPF) chicken embryos. The virus produced multifocal areas of epithelial hyperplasia along with foci of inflammation in the CAM, and rare cells contained small eosinophilic intracytoplasmic bodies. Chickens inoculated with the isolated virus in the feather follicle of the leg did not develop significant lesions. Nucleotide sequence comparison of a PCR-amplified 4.5 kb HindIII fragment of the genome of flamingo poxvirus (FlPV) revealed very high homology (99.7%) with condor poxvirus (CPV), followed by approximately 92% similarity with canary poxvirus (CNPV) and Hawaiian goose poxvirus (HGPV), but less similarity (approximately 69%) to fowl poxvirus (FPV), the type species of the genus Avipoxvirus of family Poxviridae. As in the cases with CPV, CNPV, and HGPV, genetic analysis of FlPV revealed an absence of three corresponding FPV open reading frames (ORF199, 200, and 202) and an absence of any reticuloendotheliosis virus (REV) sequences in this region. There are only nine nucleotide substitutions observed between FlPV and CPV in the 4.5 kb fragment; those were clustered in the ORF201 region, which in FPV genome is a site for integration of REV sequences. Phylogenetic analysis of the predicted amino acid sequences of the ORF201-coded hypothetical protein demonstrated FlPV to be more closely related to CPV, as well as to CNPV and HGPV, than to FPV. 相似文献
14.
20只外购的KM和BALB/c小鼠发生了酷似鼠脱脚病的典型症状，取病鼠的肝、脾、肾、淋巴结和脑作组织学检查，发现其病理变化呈痘病毒感染的特征性改变，组织镁浆液接种传代的BHK细胞，可发生明显的CPE变化，经电镜超薄切征及负染检查发现大量的痘病毒颗粒，粒子的中心为DNA核酸和蛋白质组成的拟核，在衣壳的外面有囊膜包裹，对鸡红细胞均呈阳性凝集反应（HA为1：16），用小鼠痘病毒ELISA试剂盒检测病鼠血清呈阳性反应，用病毒悬液免疫接种KM和BALB／C小鼠后，发现前者感染性强，发病快，症状典型，以急性为主，并迅速出现死亡。后者对鼠痘抵抗力强，一般呈陷性感染，特征性症状出现较缓慢，但后者一旦施加实验因素动物则出现症状，也可迅速死亡。实验证明，该群小鼠患的传染病病原为鼠痘病毒。 相似文献
15.
Avian poxvirus was isolated from nodules on the heads and conjunctiva of two 3-to-4-wk-old ostrich chicks. The ostriches from which poxvirus was isolated had been placed on premises where turkeys that had shown evidence of poxvirus infection had been raised earlier. Microscopically, the nodules from the ostriches were composed of proliferating and hypertrophic epithelial cells that formed large fronds. Most of the hypertrophic epithelial cells contained large eosinophilic intracytoplasmic inclusion bodies characteristic of poxvirus. Characterization of the avian poxvirus isolated from the cutaneous lesions in ostriches was based on western blotting of virus antigen, restriction fragment length polymorphism of genomic DNA, pathogenesis, and cross-protection studies in chickens. Antigenic and genetic studies did not reveal any significant difference between the poxvirus isolated from ostriches (PVO) and fowl poxvirus (FPV). Further, susceptible chickens immunized with the PVO were protected when challenged with a virulent strain of FPV. Thus, the poxvirus isolated from ostriches had similar antigenic, genetic, and biological properties to FPV. 相似文献
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An avian poxvirus from cutaneous lesions in a Hawaiian goose (Branta sandvicensis) was characterized in this study. The virus was isolated by inoculation onto the chorioallantoic membranes (CAMs) of developing chicken embryos. Cytoplasmic inclusion bodies were observed on histopathological examination of CAM lesions. Western blotting analysis using polyclonal antiserum against fowl poxvirus (FWPV) showed differences from FWPV, but a similar antigenic profile between Hawaiian goosepox (HGP) isolate and two previous Hawaiian poxvirus isolates were observed. Still three avian poxviruses from Hawaiian birds showed distinguishable reaction in approximately 27, 34, 35, and 81 kDa proteins when polyclonal antibodies against the Hawaiian poxvirus isolate (Alala/lanakila) were used. Restriction fragment length polymorphisms (RFLP) of DNA of this isolate also showed differences from those of FWPV and previous avianpox isolates from Hawaiian forest birds. While nucleotide sequences of a 5.3-kb PstI-HindIII fragment of the genome of HGP isolate revealed very high homology (99% identities) with Canary poxvirus (CNPV) ORF266-274, and like CNPV, homologs of three FWPV ORFs (199, 200, and 202) including any reticuloendotheliosis virus (REV) sequences are not present in the genome of HGP isolate. 相似文献
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W Herbst H Krauss 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1989,36(6):477-479
From liver, lung, and kidney of a dead sparrow with signs of conjunctivitis a cytopathic agent was isolated in chicken embryo cell culture which was identified as a poxvirus by electron microscopy. From its growth characteristics in avian and mammalian cell culture it was concluded to belong to the genus avipox virus. 相似文献
19.
Y. S. Chung D.V.M. Ph.D. P. B. Spradbrow B.V.Sc. Ph.D 《Australian veterinary journal》1977,53(7):334-336
An avian poxvirus was isolated from a black-backed magpie in Queensland. The virus produced pocks on the chorio-allantoic membrane of chicken embryos and lesions on the skin of chickens. Comparison by passive haemagglutination test and neutralisation test indicated that the virus was related more closely to pigeonpox virus than to fowlpox virus. 相似文献
20.
Lamien CE Le Goff C Silber R Wallace DB Gulyaz V Tuppurainen E Madani H Caufour P Adam T El Harrak M Luckins AG Albina E Diallo A 《Veterinary microbiology》2011,149(1-2):30-39
Sheep poxvirus (SPPV), Goat poxvirus (GTPV) and Lumpy skin disease virus (LSDV) are Capripoxviruses (CaPVs) responsible for causing severe poxvirus disease in sheep, goats and cattle, respectively. Serological differentiation of CaPVs is not possible and strain identification has relied on the implicitly accepted hypothesis that the viruses show well defined host specificity. However, it is now known that cross infections can occur and authentication of identity based on the host animal species from which the strain was first isolated, is not valid and should be replaced with molecular techniques to allow unequivocal strain differentiation. To identify a diagnostic target for strain genotyping, the CaPV homologue of the Vaccinia virus E4L gene which encodes the 30 kDa DNA-dependent RNA polymerase subunit, RPO30 was analyzed. Forty-six isolates from different hosts and geographical origins were included. Most CaPVs fit into one of the three different groups according to their host origins: the SPPV, the GTPV and the LSDV group. A unique 21-nucleotide deletion was found in all SPPV isolates which was exploited to develop a RPO30-based classical PCR test to differentiate SPPV from GTPV that will allow rapid differential diagnosis of disease during CaPV outbreaks in small ruminants. 相似文献