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《河南畜牧兽医(综合版)》1999,(12)
鸡新城疫病毒单抗及强毒感染快速诊断技术新城疫强毒感染的快速诊断成为多年未解决的世界性难题。本项目以３种不同毒力型的新城疫病毒为免疫原研制出多种单抗,从中筛选出两株单抗,建立单抗夹心聚乙二醇—酶联免疫吸附试验,可用于检测现场样品中存在的微量新城疫病病毒抗原,可以替代鉴定新城疫病毒毒力型所用的费时、费钱的常规生物试验。研制出的快速检测新城疫强毒感染的单抗诊断试剂盒已在６省市４００多个鸡场几千万只鸡的群体中应用,挽回经济损失近亿元。该项目建立的快速检测新城疫强毒的单抗夹心聚乙二醇—酶联免疫吸附试验在程… 相似文献
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为建立一种快速的新城疫病毒抗体检测方法,本研究以原核表达的重组HN蛋白作为诊断抗原,建立了检测新城疫病毒抗体的间接ELISA诊断方法.该方法检测AIV(H9亚型)、IBV、IBDV、EDSV 4种常见禽病病原的阳性血清均为阴性;检测灵敏度为1:12 800;批内重复性试验、批间重复性试验的变异系数分别小于5％和10％;与血凝抑制试验(HI)符合率为96.1％.本研究建立的NDV HN间接ELISA检测方法具有良好的特异性、敏感性和重复性,为NDV的抗体检测及流行病学调查等快速诊断提供一种技术手段. 相似文献
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乳胶凝集试验检测鸡新城疫病毒血清抗体的研究 总被引：12，自引：1，他引：12
用硫酸铵沉淀、透析浓缩的鸡新城疫病毒（ＮＤＶ）抗原致敏乳胶,然后所鸡新城疫阳性血清进行方阵滴定,选择出最佳致敏条件并制成新城疫病毒乳胶抗原,由此建立超乳胶凝集试验（ＬＡＴ）,用来检测新城疫血清抗体;用所建立的乳胶凝集试验（ＬＡＴ）和血凝抑制试验（ＨＩ）同步检测６９份鸡血清,结果乳胶凝集试验阳性６２份,阴性７份;血凝抑制试验阳性６６份,阴性３份,两者的总符合率为９４．２％（６５／６９）,且两者的检出率基本一致。结果表明,乳胶凝集试验具有操作简便、快速、敏感性高、特异性强且可用于现场检测等优点,是一种适合基层单位用来检测鸡新城疫病毒血清抗体的新方法。 相似文献
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为了对目前流行的鸭新城疫进行快速诊断,应用蔗糖密度梯度离心纯化后的鸭新城疫病毒免疫6周龄雌性Balb/c小鼠,取免疫后小鼠脾细胞与骨髓瘤细胞SP2/0融合.采用ELISA方法筛选阳性细胞克隆,经3次亚克隆后获得一株能稳定分泌鸭新城疫病毒特异性单克隆抗体的细胞株2C1.以PEG纯化后的2C1腹水为捕获抗体,辛酸-硫酸铵纯化的兔抗鸭新城疫病毒多克隆抗体为第二抗体,HRP标记的羊抗兔抗体作为酶标抗体,建立了检测鸭新城疫病毒抗原的双抗体夹心ELISA方法.结果表明,当2C1包被量为4μg/孔,鸭新城疫病毒1:10稀释,兔抗鸭新城疫病毒多抗作用量为0.1 μg/孔,HRP标记的羊抗兔二抗1:10 000倍稀释,每次作用时间为1h时,P/N比值最高,且阳性OD值最接近1.0.试验表明,该方法特异性好,不与IBV、ARV、GPV、DPV、DHV尿囊液及H5、H9标准阳性抗原发生反应;可以检测出0.2 μg纯化病毒,比传统的HA试验的敏感性至少高64倍;批间和批内变异系数均小于10％.本研究建立的双抗体夹心ELISA方法为鸭新城疫病毒的快速诊断奠定了基础. 相似文献
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ELISA检测鸡新城疫病毒特异性IgM抗体的研究 总被引：3，自引：0，他引：3
以新城疫病毒单克隆抗体包被板,用10％小牛血清-PBS封闭后,捕获尿囊液中的新城疫病毒作固相抗原.在此板上应用酶标抗鸡IgM单克隆抗体进行间接ELISA试验检侧鸡血清中新城疫病毒的特异性IgM抗体.试验证明该方法特异性强、敏惑性高.兔抗新城疫病毒阳性血清可特异性阻断反应,将新城疫病IgM阳性血清用2-ME处理可使ELISA反应呈阴性,与禽源多杀性巴氏杆菌鸡IgM阳性血清、鸡传染性支气管炎病毒IgM阳性血清无交叉反应.该试验可检测到La Sota免疫后3天鸡血清中的特异性IgM,对鸡新城疫病毒IgM阳性血清的检测效价可达1:320以上,并可检测到临床新城疫病鸡血清中的特异性IgM抗体. 相似文献
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《中国家禽》2017,(14)
为了掌握邵武市鸭新城疫病毒感染情况,2016年秋季采用鸡新城疫病毒抗原对该市19个乡镇、街道办事处随机抽样采集的散养鸭和规模鸭场血清样品,进行鸭新城疫病毒非免疫抗体检测,共检测血清样品177份,其中新城疫病毒抗体阳性的鸭血清样品93份,阳性率高达52.54%。部分散户鸭新城疫病毒抗体阳性率高达100%,散户鸭新城疫病毒平均抗体阳性率为60%,规模鸭场平均抗体阳性率为45%,这可能与鸡鸭混养的养殖模式有关。从检测结果还发现,非免疫新城疫病毒抗体的阳性率与鸭的品种有关,易感性依次为番鸭麻鸭北京鸭。总体显示鸭新城疫病毒的感染情况虽然高,但只感染不发病,结果对水禽新城疫的防控有一定的指导意义。 相似文献
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Xiaoyu Deng Yanlong Cong Renfu Yin Guilian Yang Chan Ding Shengqing Yu Xiufan Liu Chunfeng Wang Zhuang Ding 《Journal of veterinary science (Suw?n-si, Korea)》2014,15(4):511-517
Characteristic clinical manifestations of Newcastle disease include leukopenia and immunosuppression. Peripheral blood mononuclear cells (PBMCs) are the main targets of Newcastle disease virus (NDV) infection. To survey changes in proteomic expression in chicken PBMCs following NDV infection, PBMC proteins from 30 chickens were separated using two-dimensional electrophoresis (2-DE) and subjected to mass spectrometry analysis. Quantitative intensity analysis showed that the expression of 78 proteins increased more than two-fold. Thirty-five proteins exhibited consistent changes in expression and 13 were identified as unique proteins by matrix assisted laser desorption ionization-time of flight mass spectrometer/mass spectrometer including three that were down-regulated and 10 that were up-regulated. These proteins were sorted into five groups based on function: macromolecular biosynthesis, cytoskeleton organization, metabolism, stress responses, and signal transduction. Furthermore, Western blot analysis confirmed the down-regulation of integrin-linked kinase expression and up-regulation of lamin A production. These data provide insight into the in vivo response of target cells to NDV infection at the molecular level. Additionally, results from this study have helped elucidate the molecular pathogenesis of NDV and may facilitate the development of new antiviral therapies as well as innovative diagnostic methods. 相似文献
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几种主要禽疫病诊断基因芯片的制备及初步应用 总被引：5，自引：2，他引：5
进行了几种主要禽疫病诊断基因芯片制备及其初步应用研究。试验分别设计和克隆鉴定了NDV、IBV、AIV和IBDV的靶基因重组质粒。以克隆的靶基因重组质粒为模板。分别进行PCR扩增制备靶基因并纯化，以基因芯片点样仪将制备的靶基因点制在氨基化的基片上，经干燥、水合、紫外线交联和洗涤后，成功制备了NDV-IBV-AIV-IBDV诊断基因芯片。试验应用CY3荧光标记制备的探针进行芯片的检验，结果表明制备的NDV-IBV-AIV-IBDV诊断基因芯片质量好，可对NDV、IBV、AIV和IBDV进行诊断检测，具有检测灵敏性好，特异性高和芯片可重复检测的优点。试验对30个临床样品进行初步应用检测，结果表明该诊断基因芯片技术与RT—PCR检测技术检出率基本一致，并具有同步诊断检测多种疫病的优点。 相似文献
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During November 2002, six double-crested cormorants (DCCs; Phalacrocorax auritus) were found moribund in Big Pine Key, FL, exhibiting clinical signs indicative of neurologic disease. Postmortem diagnostic evaluations were performed on two adult birds. Virulent Newcastle disease virus (NDV) was isolated from a cloacal swab from cormorant 1. West Nile virus (WNV) was isolated from the brain and lung of cormorant 2. Nucleotide sequence analysis of a portion of the fusion (F) protein gene of the NDV cormorant isolate revealed it shared a 100% deduced amino acid identity with only two viruses: the 1992 epizootic cormorant isolate from Minnesota and the 1992 turkey isolate from North Dakota. The epidemiologic significance of the recognition of virulent NDV on cormorant wintering grounds during a nonepizootic period, in addition to the potential implications of the concurrent isolation of NDV and WNV from cormorants, is discussed. 相似文献
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A trial was conducted to evaluate the antiviral activity and immunomodulatory effect of B-Caryophyllene (BCP) using NDV as a viral model. First, an in ovo experiment was conducted to estimate the antiviral mechanism of BCP. Next, an in vivo experiment was designed to confirm its antiviral efficacy as well as its immunomodulatory and growth promoting ability. According to the in ovo experiment, BCP possesses antiviral influence up to 61.7% when treated before or during NDV infection. Oral supplementation of chickens with two doses of BCP (200 and 400 μg/bird) resulted in a significant increase in the NDV HI-Ab responses and a significant increase in interferon-α signaling cytokines. These obvious immunomodulatory effects improved the bird clinical protection against virulent NDV challenge. To conclude, we introduced a new compound for the poultry industry sector that has antiviral and immunostimulant properties when supplemented orally before or during NDV infection. 相似文献
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An approach that possesses high specificity and broad applicability was used to obtain a DNA probe with potential diagnostic value. By utilizing synthesized oligonucleotide DNA, termed NDV probe, all 14 strains of NDV tested under high-stringency conditions were recognized in a slot-blot hybridization assay. The sequence of the NDV probe was generated from a highly conserved region of the NDV genome. No hybridization was observed with RNA isolated from other avian viruses, including avian influenza, infectious bursal disease, and infectious bronchitis. The specificity inherent in using an oligonucleotide probe offers advantages over probes obtained from cloned DNA fragments. 相似文献
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The interference phenomenon of infectious bronchitis virus (IBV) with growth of Newcastle disease virus (NDV) in embryonating chicken eggs (ECE) was used as a diagnostic method. Fifteen field isolates obtained from presumptively infectious-bronchitis-affected chickens were analyzed by the IBV-NDV interference test. Eight isolates were capable of interfering with the growth of the La Sota strain of NDV, as measured by hemagglutination (HA) activity when IBV was inoculated 10 hr before NDV into ECE. The interference was considered specific for IBV, because it could be eliminated by adding homologous anti-IBV serum. The sensibility of this method could be demonstrated, because in some cases low-passage levels of IBV isolates showing HA interference ability were not capable of producing lesions in ECE. Furthermore, serologically negative IBV samples did not interfere with NDV growth. From these results, the IBV-NDV interference test appears to be a potential diagnostic alternative for identifying IBV field isolates. 相似文献
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FC‐16 Clinical and immunological effects of Newcastle disease virus vaccine on bovine papillomatosis
Newcastle disease virus (NDV) has antineoplastic and immunostimulatory properties, and it is currently being clinically tested in anticancer therapy. In order to analyse the immunostimulatory effects of NDV on bovine papillomatosis, we inoculated 14 cows subcutaneously with an attenuated vaccine containing the LaSota strain of NDV (LS‐NDV). Four cows with papillomatosis served as controls. Serum samples were collected from each animal 1 h preinoculation and 7 and 21 days postinoculation. In inoculated cows on days 7 and 21, the mean antibody titres were log2 2.43 ± 0.92 and 5.57 ± 0.72, respectively, by haemagglutination inhibition, and the mean levels of TNFα were 5.80 ± 4.19 and 5.39 ± 2.66 ng/mL, respectively, by WEHI‐164 cytotoxicity assay. Significant differences between inoculated and control animals were evident for antibody titres on day 21 and clinical scores on day 60. A correlation was evident between the TNFα activities and clinical scores on day 21. The clinical observations at day 60 showed that the papillomas in five cows had completely resolved (36%), one animal had no alterations on clinical appearance of the tumour (7%), and papillomas in eight cows had regressed (57%). In conclusion, these results demonstrated that inoculation of LS‐NDV vaccine stimulates an antibody response and a limited increase in TNFα activity and may enhance clinical recovery in bovine papillomatosis. Funding: Scientific Research Council of Akdeniz University. 相似文献
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试验旨在观察鸡传染性支气管炎病毒（infectious bronchitis virus,IBV）和鸡新城疫病毒（newcastle disease virus,NDV）之间的增殖干扰现象,分析在免疫过程中两种疫苗存在的免疫干扰,为确定疫苗的免疫程序提供依据。采用完全随机试验设计,选取10日龄鸡胚尿囊腔接种不同浓度的IBV、NDV以及不同浓度混合的IBV和NDV,收集尿囊液测定其血凝效价（HA）;用不同浓度的IBV、NDV以及不同浓度混合的IBV和NDV分别免疫BALB/c小鼠及SPF雏鸡,收集血清,间接酶联免疫吸附法（ELISA）和血凝抑制（HI）试验测定IBV和NDV的抗体效价及抑制效价。结果表明,同胚培养时,无论先接种IBV后接种NDV还是先接种NDV后接种IBV或IBV和NDV同时接种,IBV对NDV均有干扰作用,而NDV对IBV没有干扰作用;接种IBV和NDV的小鼠,其产生IBV和NDV的抗体效价均低于单独免疫组,免疫的次序及免疫间隔时间对IBV和NDV的抗体效价均有影响,IBV对NDV的免疫效果具有较大的干扰作用。不同针混合免疫方式能提高IBV和NDV的抗体效价;接种IBV和NDV的雏鸡,免疫次序及免疫相隔时间对IBV和NDV的抗体效价均有影响,IBV对NDV的免疫效果具有较大的干扰作用;雏鸡和小鼠免疫血清HI试验数据表明,免疫次序及免疫间隔时间对IBV和NDV抗体的产生均有影响,但对NDV抗体的产生影响更明显,随着免疫间隔时间的增加,IBV和NDV的抗体水平呈增加趋势,雏鸡比小鼠更能敏感地反映出IBV对NDV的免疫干扰作用。混合注射时IBV和NDV的抗体水平均降低,IBV对NDV的免疫效果有干扰作用。因此,在同胚培养、小鼠及雏鸡的免疫中,IBV对NDV有干扰作用,而NDV对IBV无干扰作用。混合接种时IBV和NDV抗体效价均下降,IBV对NDV的免疫效果有干扰作用。免疫次序及免疫间隔时间对IBV和NDV抗体的产生均有影响,但随着免疫间隔的增加,IBV和NDV的抗体水平呈增加趋势。 相似文献