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根据禽网状内皮组织增生症病毒SNV株的前病毒基因组cDNA序列设计并合成1对引物,以pPB101质粒为模板,通过PCR扩增获得该病毒p30基因片段。将PCR产物克隆入表达载体pCold-HF中,构建的原核表达质粒pCold-p30转化BL21（DE3）菌,经IPTG诱导,p30蛋白呈现可溶性表达。用表达产物免疫6周龄BALB/c小鼠,制备p30抗体,经Westernblot和间接免疫荧光法检测,结果发现,原核表达的SNV株p30蛋白具有良好的免疫原性。 相似文献
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本文根据禽痘病毒（FPV）基因组中禽网状内皮组织增生症病毒（REV）常见整合区域的序列设计合成一对引物，对国外一株FPV疫苗进行了PCR扩增，结果扩增出一条1478bp的片段，经过测序和序列分析表明该片段为REV和FPV的整合序列，FPV序列中整合了大小为193bp的REV长末端重复序列，进一步比较显示该序列与国外REV毒株APC566、713和SNV的同源性分别为：99．6％、98.0％和87．3％，与我国REV分离株HA9901的同源性只有83．6％。同时该REV整合序列与国外已经测定的FPV毒株AJ581527、AY255633、AF198100、AF006064和AF246698比较时发现，这些毒株中含有的REV序列不仅完全一致，而且REV序列的插入位点也完全相同。这说明国外FPV疫苗中不仅含有REV整合序列，而且已经存在多年了。 相似文献
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本试验采集广东某鸡场疑似禽网状内皮组织增生症患病鸡病料并进行处理,经接种DF1细胞、聚合酶链式反应（PCR）及特异性间接免疫荧光试验（IFA）,分离鉴定出1株禽网状内皮组织增生症病毒（REV）,命名为 GD1210。依据已发表的REV前病毒全基因组序列设计并合成6对引物,采用分段扩增的方法完成了分离株的全基因组序列测定。结果显示,该分离株基因组序列全长8443 bp。将该分离株的基因序列与国内外各参考株相应序列进行同源性比对分析,与SNV株亲缘关系最近,核苷酸同源性最高。将该分离株感染1日龄SPF鸡以鉴定其致病性,结果显示该分离株使SPF鸡产生严重的免疫抑制作用。 相似文献
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《中国预防兽医学报》2016,(11)
为比较分析近年来禽网状内皮组织增生症病毒(REV)的基因组变异情况,本研究对分离自某父母代肉种鸡场患病鸡的REV LN1201株进行PCR扩增和克隆测序,获得了LN1201株的前病毒全基因组序列,并将其与不同参考株基因组进行了比较分析。结果显示,LN1201株前病毒基因组全长8 438 bp,具有典型复制完整型逆转录病毒的基因组结构。与参考株相比,LN1201与国内分离株HLJR0901和疫苗污染病毒株MD-2同源性较高,但其U3区与不同病毒株的同源性相对较低,并且在U3区出现了罕见的14 bp碱基插入,显示该病毒株可能为LTR发生重组的病毒。本研究有助于了解我国鸡群中REV的基因组特性,提示有必要继续关注REV诱发的肿瘤及其基因变异。 相似文献
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我国鸡痘病毒野毒重组株的流行初探 总被引：1，自引：1，他引：0
从山东、河南等省不同的鸡痘发病地区分离和鉴定了20株鸡痘病毒野毒株,并分别根据禽网状内皮组织增生症病毒（REV）和鸡痘病毒（FPV）的基因组设计合成了1对上游源自REV基因组和下游源自FPV基因组的引物。随机从20株FPV野毒株中挑选7株接种鸡胚成纤维细胞,提取细胞DNA作为模板,利用这对引物进行PCR反应,结果均扩增到了779bp大小的目的片段。将其中1个片段测序发现此片段为REV和FPV的整合序列,其中REV序列426个碱基,FPV序列为304个碱基。结果表明,我国FPV野毒中已整合入REV的基因序列,而且这种整合株所占比例相当高,已成为主要的流行毒株。 相似文献
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为了快速、准确检测禽源生物制品中的禽网状内皮组织增生症病毒(REV)和禽白血病病毒(ALV),根据GenBank中登录的REV参考毒株LTR基因序列和ALV P27基因序列,设计合成2对特异性引物。在建立各病毒单项PCR检测技术的基础上,优化反应条件,建立2种病毒的双重PCR检测方法。结果:可同时扩增REV的467 bp和ALV的675 bp的特异性片段,而对其他5种禽病病原的PCR扩增结果均为阴性。敏感性测定结果表明,该双重PCR技术能检出10 pg的ALV和10 pg的REV模板。用双重PCR技术与REV的间接免疫荧光法及ALV ELISA对245份疫苗株检测,进行同步比较,结果总符合率为100%。表明建立的双重PCR检测方法,具有特异、快速、准确的特点,对这2种外源病毒的同时检测,显示出了较好的可行性。 相似文献
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禽网状内皮组织增生症病毒囊膜糖蛋白gp90的原核表达 总被引：9，自引：0，他引：9
根据禽网状内皮组织增生症病毒（REV）SNV株的前病毒基因组cDNA序列，设计并合成1对引物，以SNV株前病毒CDNA全基因组克隆为模板，通过PCR技术，扩增出该病毒囊膜糖蛋白（env)gp90基因部分片段。将PCR产物按正确的阅读框架定向克隆进pGEX-5X-3载体中谷胱甘肽-S-转移酶（GST）的下游，将重组质粒转化进宿主菌BL21中，在1.0mmol/L IPTG(37℃）诱导下，gp90基因部分片段以融合蛋白的形式获得了良好的表达。表达产物经聚丙烯酰胺凝胶电泳鉴定，确定其表达的融合蛋白相对分子质量为64000。将表达产物从凝胶中回收后免疫小鼠，所得的抗血清可与REV野毒株感染的鸡胚成纤维细胞（CEF）在免疫荧光试验中起反应。由此表明，体外表达的SNV株gp90-GST融合蛋白保留了天然蛋白所具有的抗原性。 相似文献
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鸡传染性贫血病毒与禽网状内皮增生症病毒二重PCR检测方法的建立 总被引：3，自引：3，他引：0
当前养鸡业中鸡传染性贫血病毒、禽网状内皮增生症病毒混合感染的发生率日益增高,为提高CIAV、REV检测效率,根据GenBank上登录的CIAV全基因序列和REV的LTR序列分别设计了2对引物,建立了REV、CIAV二重PCR检测方法,另外还比较了2种不同的DNA提取方法。结果显示,该方法扩增目的条带清晰、敏感性高、特异性好。应用建立的方法对临床7份病料样品检测,检出5份CIAV、REV阳性,表明建立的二重PCR方法能有效用于CIAV、REV混合感染的临床诊断。 相似文献
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Pathogenesis of reticuloendotheliosis virus infection in ducks 总被引：1，自引：0，他引：1
A new isolate of reticuloendotheliosis virus (REV) from ducks, RU-1, was used to experimentally infect white pekin ducks. Embryonal or neonatal infection usually resulted in persistent viremias, no REV antibody development, and inability to mount antibody responses against bovine serum albumin (BSA) or sheep red blood cells (SRBC). In contrast, infection at 21 days of age resulted in transient viremias, which terminated coincident with REV antibody development. These ducks remained persistently infected, however, based on virus isolations from peripheral blood lymphocytes. Ducks infected at that age were immunologically competent against BSA and SRBC unless they had been embryonally bursectomized, in which case they behaved virologically and immunologically like those infected at an early age. Bursectomy by itself did not prevent responses against the antigens. Total mortality during 4- or 6-month experimental periods ranged from 80 to 100% in REV-infected groups, regardless of age at infection, route of exposure, or whether the ducks were intact or bursectomized. Most deaths were from non-neoplastic conditions (stunting, bacterial infections), but 17 of 69 (25%) infected ducks developed a variety of neoplasms, including lymphosarcomas, histiocytic sarcomas, and spindle-cell sarcomas. 相似文献
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Epizootiological studies were conducted on a commercial turkey breeder flock naturally infected with nondefective reticuloendotheliosis (RE) virus. Although RE virus was isolated from 27 (46%) of the 59 hens studied, only 4 of the 45 hens tested transmitted RE virus to progeny during a 6-week observation period and the overall transmission rate was 1.8%. The transmitter hens were of two types: three hens were consistently viremic and antigenemic and lacked antibody; one hen was viremic but lacked detectable viral antigen and possessed antibody. Toms appeared to play no role in vertical transmission of the infection. Of several tests evaluated for detection of transmitter hens, the direct enzyme-linked immunosorbent assay on albumen was probably best, since it detected three of four transmitter hens, detected relatively few nontransmitter hens, and had the best consistency of any test. No significant differences in hatchability were found between eggs from viremic and non-viremic hens. These findings can be utilized in the development of programs for eradication of RE virus from turkey breeding flocks. 相似文献
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Six broiler breeder flocks from two companies in Mississippi were tested at intervals for reticuloendotheliosis (RE) virus infection. Virus was isolated and antibody demonstrated in all six flocks. Infection was first detected at ages ranging from 13 to 47 weeks. Studies showed that neither congenital transmission from grandparent flocks nor treatment with contaminated vaccines was a likely source of infection; thus, exposure to RE virus was assumed to come from the environment. Virus was isolated from litter samples from two of the flocks, but no specific environmental infection source was identified. Infection rates of flocks differed between the two companies. Although adequate controls were lacking, no performance problems due to RE virus infection were apparent in breeder or broiler progeny flocks. However, the RE viruses isolated from these flocks were immunosuppressive and oncogenic when inoculated into day-old chicks. A moderate (3-16%) incidence of neoplasms was induced by contact exposure to these field isolates in the laboratory. 相似文献
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商品代肉鸡群中禽网状内皮组织增生病病毒和马立克氏病病毒的混合感染 总被引：6，自引：0，他引：6
山东兖州某商品代肉鸡场的 2 0日龄肉鸡开始出现大批死亡 ,剖检发现肝脏、脾脏、肾脏和心脏等主要脏器均出现大小不等的肿瘤结节 ,采用细胞培养和间接荧光抗体试验的方法从这些病料中同时分离到了禽网状内皮组织增生病病毒 (REV)和马立克氏病病毒 (MDV) ,通过组织病理学检查和斑点杂交的方法表明该鸡场的疫病为REV和MDV共感染引起的肿瘤病。这是国内从商品代肉鸡中同时分离到REV和MDV的首次报道 相似文献
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鸡肿瘤病料中马立克氏病病毒和禽网状内皮组织增生症病毒共感染的研究 总被引：24，自引：1，他引：23
采用细胞培养、间接荧光抗体试验(IFA)、聚合酶链式反应(PCR)和斑点杂交(Dotblot)的方法从我国不同地区发生肿瘤的病料中同时进行MDV和REV的分离和鉴定。在分离到的13株MDV野毒株中，有4株培养物既能在IFA中与REV的单抗反应，又可以用PCR扩增出REV的LTR；另有4株培养物能扩增出REV的LTR，但在IFA中却不与REV的单抗反应。结果表明我国MD肿瘤中存在着REV的共感染，且我国MDV某些野毒株的基因组中有可能已经整合进了REV的LTR序列。 相似文献
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为研究鸡源网状内皮组织增殖病病毒(REV)对HBK无特定病原体(SPF)鸭的致病性,利用鸡源REV现地分离株REV-HN,分别接种鸭胚成纤维细胞(DEF)、鸭胚(尿囊腔和卵黄囊腔接种)和雏鸭(口腔接种),试验设空白对照组,雏鸭还设有同居感染组.接种REV后不同时间收获DEF和胚体,及雏鸭的外周血淋巴细胞(PBL)和免疫器官,提取基因组DNA,通过RT-PCR和PCR方法检测REV前病毒env基因.结果在DEF中未检测到特异性的env基因片段,而在卵黄囊接种后72 h鸭胚胚体和试验感染的雏鸭PBL中检测到.接种后30 d,在1羽感染REV的雏鸭肾、胸腺和法氏囊中检测到REV前病毒.本研究为利用SPF鸭研究REV提供了依据. 相似文献
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P S Paul K A Pomeroy C C Muscoplat B S Pomeroy P S Sarma 《American journal of veterinary research》1977,38(3):311-315
Reticuloendotheliosis (RE) virus strains MN81 and MN67 isolated from epiornithics of RE in turkeys were partially characterized. Strains MN81 and MN67 replicated in chicken embryo fibroblast,duck embryo fibroblast and turkey embryo-fibroblast cultures and produced syncytial cytopathic effects in duck embryo fibroblast and turkey embryo fibroblast cultures. The virions of MN81 and MN67 measured approximately 100 nm in diameter, resembled RE virus strain T, and could be distinguished from avian leukosis viruses morphologically. The buoyant density of strain MN81 was found to be 1.15 g/cm3 in sucrose gradients. Strains MN81 and MN67 were inactivated by heat, acid pH, ether, and chloroform treatments. These strains were serologically unrelated to avian leukosis virus but were related to RE virus strains T, CS, DIA, and SN. 相似文献
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禽网状内皮组织增生病病毒分子生物学特性与免疫抑制 总被引：1，自引：0，他引：1
禽网状内皮组织增生病病毒为反转录病毒科禽类C型反转录病毒.禽网状内皮组织增生病病毒感染后主要引起感染家禽免疫功能抑制和慢性肿瘤,该病的控制对养禽业的健康发展和生物制品质量的提高具有重大影响.文章对禽网状内皮组织增生病病毒的生物学特性与引起免疫抑制机制的研究新进展进行了简要综述. 相似文献