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将减蛋综合征(EDS-76)H株和新城疫(ND)La Sota弱毒株分别接种鸭胚和鸡胚,收获胚液(HA价分别达2~(15)和2~(11)以上),福尔马林灭活后配制成EDS-ND二联油佐剂灭活苗。经无菌与安全性检验,免疫效力测定和田间应用结果表明,该苗安全性良好,于蛋鸡开产前1个月左右皮下或肌肉注射0.5ml/羽,免疫后21天HI抗体效价均达1:2~(10)以上,免疫期达6个月以上。 相似文献
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正产蛋下降综合征是一种由禽腺病毒引起的,以产蛋量下降为特征的禽类传染病。病鸡无明显症状,而以产蛋量骤然下降、蛋壳异常(产软壳蛋、薄壳蛋)、产畸形蛋、蛋质低劣和蛋壳颜色变淡为特征。1病原产蛋下降综合征病毒属于禽腺病毒,在7~10日龄鸭胚中生长良好,可使鸭胚致死,其尿囊液具有很 相似文献
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用鸡减蛋综合征(EDS)BH_9i毒株接种鸭胚,收获鸭胚尿囊液,用一定浓度福尔马林灭活后加入吐温80制成水相,司班80和硬脂酸铝加入白油中制成油相,二者按一定比例乳化后制成EDS油乳剂灭活苗,该苗接种开产前母鸡,可使接种鸡获得对EDS强毒攻击坚强的抵抗力,该苗安全、稳定,4℃保存可达12个月以上。 相似文献
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两株鸡减蛋综合症病毒在鸭胚中增殖规律的观察毛春生，卢中华，崔保安，李季枝，陈丽颖，王英汉（河南农业大学牧医系）减蛋综合征（ＥＤＳ）是由禽腺病毒引起的鸡的传染病，该病以病鸡产蛋减少和蛋壳形成不良为特征。目前世界上许多国家和地区都有流行［１，２］。该病已... 相似文献
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用鸡 减蛋综合征（ＥＤＳ）ＢＨ９１毒株接种鸭胚，收获鸭胚尿囊液，用一定浓度福尔马林灭活后加入吐温８０制成水相，司班８０和硬脂酸铝加入白油中制成油相，二者按一定比例乳化后制成ＥＤＳ油乳剂灭活苗，该苗接种开产前母鸡，可使接种鸡获得对ＥＤＳ强毒攻击坚强的抵抗力，该苗安全，稳定，４℃保存可达１２个月以上。 相似文献
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鸡新城疫-传染性支气管炎-减蛋综合征三联油乳剂苗的研制及应用 总被引：1，自引：0，他引：1
用新城疫病毒（克隆H88株。）、传染性支气管炎病毒（JP株和H52株）和减蛋综合征病毒（H91株）分别接种于鸡胚和鸭胚,并收取其含毒鸡、鸭胚尿囊液,用一定浓度甲醛溶液灭活,按一定比例混匀,以7号白油为佐剂制成鸡新城疫（ND）－传染性支气管炎（IB）－减蛋综合征（EDS76）三联油乳剂灭活苗（三联苗）。三联苗免疫30日龄雏鸡,免疫后7天产生免疫应答,免疫后21天保护率达100％。疫苗在4℃～8℃保存期为12个月,10℃～25℃保存期为6月,该疫苗在全省不同地区鸡群应用400余万羽份,取得满意效果。 相似文献
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为优化鸡减蛋综合征相关多联灭活疫苗的生产工艺,利用Pellicon切向流超滤系统,将制备的鸡减蛋综合征病毒液6.5倍浓缩,0.1%(V/V)甲醛灭活。将抗原液用灭菌生理盐水进行1∶8、1∶16、1∶32、1∶64、1∶128、1∶256稀释,按照常规生产工艺规程分别制备灭活疫苗,免疫21日龄SPF雏鸡。21d后翅静脉采血,HI试验检测免疫鸡的抗体水平。结果显示,免疫鸡HI抗体效价几何平均值均不低于7.0log2,符合鸡减蛋综合征灭活疫苗质量标准要求。 相似文献
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鸭黄病毒病为近年来新发的鸭病,给养鸭业带来了极大的经济损失。为了深入研究本病的防制,本试验对鸭黄病毒(duck flavivirus,DFV)进行了分离鉴定。取疑似感染DFV的病鸭病料,经细菌分离初步排除细菌感染后,应用RT-PCR检测呈现DFV阳性,处理后将其接种到鸭胚成纤维细胞(DEF)和健康鸭胚上进行病毒分离传代。结果显示,在DEF细胞上第1代48 h就开始出现CPE,随着时间的延长CPE更加明显,通常在72~96 h产生典型CPE;接种鸭胚每一代均出现死亡,且死亡时间多集中于接种后60~72 h,死亡鸭胚胚体水肿、出血、发育不良、胚肝严重出血、肿胀或斑驳样坏死等病变。将病毒DEF细胞和鸭胚分离物应用血凝试验、毒价测定、病毒中和试验、RT-PCR及人工感染试验进行检测鉴定,证明所分离到的病毒为DFV,并将其命名为DFV SDbz株。 相似文献
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鸭疱疹病毒Ⅱ型（暂定名）的分离鉴定 总被引：11，自引：3，他引：8
自1990年以来，在福建、浙江和广东等省发生一种以双翅羽毛管淤血呈紫黑色、断裂和脱落以及皮下、脏器（肝脏、胰脏）和肠道（十二指肠、直肠和盲肠）出血为主要特征的新的鸭传染病，暂定名为鸭出血症。我们对从自然感染该病典型病死番鸭脏器中获得的1株含2种病毒粒子（直径大小分别为30－40mm和80－120mm)的分离物，以Ⅰ型雏鸭病毒性肝炎标准阳性血清进行连续4代中和后接种番鸭胚，收集死亡番鸭胚胚液进行病毒纯化、负染、电镜观察和SPF鸡胚接种试验证明获得单一病毒分离株（定名为鸭出血症病毒）。该病毒为不耐酸、不耐碱、不耐热、对氯仿处理敏感、核酸类型为双股DNA的有囊膜病毒，直径为80－150nm；对番鸭胚、鹅胚、麻鸭胚、半番鸭胚、北京鸭胚和SPF鸡胚的致 死率分别为100％、100％、78．3％、60％、82．1％和0％；对10－11日龄番鸭胚的ELD50为10^-7.78/0.2ml；无血凝活性，不凝集“O”型人、鸭、鸡、鹅、家兔、小鼠，豚鼠、猪和绵羊红细胞；经血清中和试验证明该病毒与Ⅰ型雏鸭肝炎病毒、雏番鸭细小病毒、雏鹅细小病毒、鸭瘟病毒无血清学相关性。据以上结果可将该病毒确定为疱疹病毒科成员，但鉴于其与鸭瘟病毒（鸭疱疹病毒Ⅰ型）无血清学相关性，则暂定名为鸭疱疹病毒Ⅱ型，此为国内外首次报道。 相似文献
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试验旨在利用胶体金免疫层析技术建立快速检测犬血清中犬细小病毒（canine parvo virus, CPV）血凝抑制（haemagglutination inhibition, HI）抗体效价的方法,用于CPV疫苗免疫效果评价。采用双抗体夹心法,以抗CPV血凝相关抗原的单克隆抗体制备CPV抗原检测试纸条;将犬血清进行不同比例系列稀释后,分别与定量CPV抗原充分反应,滴入CPV胶体金试纸条,根据试纸条检测线（test line,T线）消失时的血清最高稀释倍数判断血清中CPV抗体的HI效价;用此方法检测86份犬血清样品,并与传统血凝抑制试验方法进行分析比较。结果显示,成功制备CPV抗原检测试纸条,确定了试纸条检测犬血清CPV-HI效价的反应条件和结果判定标准。结果表明,在检测不同稀释倍数犬血清反应后的CPV抗原时,能使试纸条T线消失时的血清最高稀释倍数与HI效价具有正相关性,犬血清最高稀释倍数乘以4即为HI效价;两种方法的符合率达90.7%。本试验初步建立了胶体金试纸条检测CPV血凝抑制效价的方法,为检测CPV-HI效价提供了一种操作简单、快速的试验方法,可用于CPV疫苗免疫效果评价。 相似文献
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MA Yong-ying SUN Ming SHEN TU Fen-qin KONG Han-jin YANG Xin-yan QIN Ya-man CHEN Xi-zhao JIA Hong 《中国畜牧兽医》2016,43(8):1983-1988
The study was aimed to use colloidal gold immune chromatography technology to establish a rapid method for detection of canine serum canine parvovirus (CPV) hemagglutination inhibition (HI) titer and CPV vaccine immunization effect assessment.Double antibody sandwich method and monoclonal antibodies of anti-CPV hemagglutination antigen were used to prepare CPV antigen test strip.Canine serum with different proportion respectively was mixed with quantitative CPV antigen for full reaction,then dropped the mixture into the CPV colloidal gold test strip,so according to the highest serum dilution ratios when the test strip line T (line T) vanishes,it was to judge CPV antibodies in serum of the HI titer.This method had been used to detect 86 canine serum samples,at the same time,analyzing and comparing it with traditional hemagglutination inhibition test method.The results showed that the CPV antigen detection test strip was successfully prepared,and the reaction conditions and results of the test strip for detecting the titer of CPV-HI in canine serum were determined.The results indicated that when detecting CPV antigen after the dilution of different ratios of canine serum,the highest serum dilution ratios when the strip line T vanished and the HI titer had positive correlation.The highest dilution ratios of canine serum multiplied by 4 was the HI titer.The results of two methods had 90.7% consistency.This experiment established the colloidal gold immune chromatography test strip for the detection of CPV-HI titers method initially.This CPV-HI detection provided a simple and fast test method for the effect evaluation of CPV vaccine immune. 相似文献
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In order to determine whether the current field strains of egg drop syndrome (EDS) 1976 viruses adapt to chickens, we compared the growth efficiency of three Japanese field strains (PA-1/79, AWI/98, Gifu/01) in chicken and duck embryo liver cells. The growth efficiency in chicken or duck embryo liver cells was almost similar in these strains. The fiber protein may carry the type-specific antigen and the hemagglutination activity, and hexon protein may contain the subgroup-specific antigenic determinants. Therefore, the fiber head and hexon loop 1 DNA domain sequences of the six Japanese field strains UPA-1/79, ME/80, 44/81, Kyoto/91, AWI/98, Gifu/01) were compared, but these DNA domains were identical among the six field strains. Our data suggested that the EDS virus was maintained without discernible changes for the last two decades in the field. 相似文献
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猪细小病毒（Porcine Parvovirus,PPV）血凝抑制试验抗原、阳性血清和阴性血清在猪细小病毒制品的效力检验中不可或缺,对猪细小病毒相关生物制品的质量控制至关重要。试验采用PPV 7909株病毒同步接种PK-15细胞制备血凝抑制试验抗原,用制备的抗原乳化后免疫豚鼠制备阳性血清,同时用未免疫的阴性豚鼠制备阴性血清。对抗原、阳性血清和阴性血清进行鉴定,结果表明,制备的PPV血凝抑制试验抗原HA效价达1:512;阳性血清HI效价达1:1024;阴性血清HI效价＜1:8,且特异性均良好。利用制备的血凝抑制试验抗原、阳性血清与不同PPV疫苗株的灭活抗原、阳性血清进行交叉反应试验,结果表明,制备的抗原与阳性血清具备良好的血清学交叉反应性,可用于PPV制品的统一评价。 相似文献
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Kang-Seuk Choi Soo-Jeong Kye Woo-Jin Jeon Mi-Ja Park Saeromi Kim Hee-Jung Seul Jun-Hun Kwon 《Journal of veterinary science (Suw?n-si, Korea)》2013,14(3):291-297
A recombinant hemagglutinin-neuraminidase (rHN) protein from Newcastle disease virus (NDV) with hemagglutination (HA) activity was expressed in Spodoptera frugiperda cells using a baculovirus expression system. The rHN protein extracted from infected cells was used as an antigen in a hemagglutination inhibition (HI) test for the detection and titration of NDV-specific antibodies present in chicken sera. The rHN antigen produced high HA titers of 213 per 25 µL, which were similar to those of the NDV antigen produced using chicken eggs, and it remained stable without significant loss of the HA activity for at least 12 weeks at 4℃. The rHN-based HI assay specifically detected NDV antibodies, but not the sera of other avian pathogens, with a specificity and sensitivity of 100% and 98.0%, respectively, in known positive and negative chicken sera (n = 430). Compared with an NDV-based HI assay, the rHN-based HI assay had a relative sensitivity and specificity of 96.1% and 95.5%, respectively, when applied to field chicken sera. The HI titers of the rHN-based HI assay were highly correlated with those in an NDV-based HI assay (r = 0.927). Overall, these results indicate that rHN protein provides a useful alternative to NDV antigen in HI assays. 相似文献
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Starick E Werner O Schirrmeier H Köllner B Riebe R Mundt E 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2006,53(8):370-375
A recombinant baculovirus (RBV) encoding the nucleoprotein (NP) of avian influenza virus (AIV) was generated and the appropriate protein was expressed in Sf9 cells. Purified recombinant NP and the NP-specific monoclonal antibody HB65 were used to establish a competitive ELISA (cELISA) system for the detection of NP-specific antibodies in sera of ducks, geese and wild birds. Tests to evaluate this method were carried out using sera of ducks experimentally infected with AIV, pre-immune duck and chicken sera, and poultry field sera, which tested negative in the haemagglutination inhibition (HI) assay, and field sera of several poultry species experimentally infected with other viruses. The evaluation of the test demonstrated a high sensitivity and specificity of this method. Tests carried out using field sera of duck and goose flocks revealed widely corresponding results obtained by HI assay and cELISA indicating that this test is applicable for flock diagnosis. Differing results were obtained for individual samples. It can be assumed that for the most part this was because of a better recognition of the conserved NP antigen by serum antibodies, although some results remained unclear. 相似文献
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Many commercial enzyme-linked immunosorbent assays (ELISAs) are unable to differentiate antibody responses to different avian influenza virus (AIV) subtypes. Developing an ELISA for specifically detecting the H5 antibody is the purpose of this study. Four monoclonal antibodies (Mabs) were raised using A/duck/Yunlin/04 (H5N2). They were confirmed as being specific to H5. Two of these antibodies showed hemagglutination inhibition (HI) activity using the HI test. Using immunodot blot assays, three Mabs recognized both Eurasian and American H5, whereas the other Mab recognized only the tested Eurasian H5 virus. When testing denatured H5 antigen, one of the Mabs lost its antigen binding activity using Western blotting. For detecting the H5 humoral response in serum, one monoclonal antibody was purified and labeled with horseradish peroxidase to set up a blocking ELISA. Chicken sera that blocked H5 Mab binding by > 29% were considered H5 antibody positive. Inhibition percentages for sera from chickens infected with other AIV subtypes, H1 to H15, were < 29%. This blocking ELISA was used for 478 field chicken serum samples. The results showed that the sensitivity and specificity of this ELISA were 98.3% (232/236) and 95.9% (232/242), respectively. This blocking ELISA could be used specifically for detecting the H5 humoral responses in chickens. 相似文献