鸡传染性支气管炎病毒血凝谱的研究

用家兔A型魏氏梭菌培养液处理的6株鸡传染性支气管炎病毒,以方阵试验分别与人O型红细胞及羊、猪、兔、鸡、鸭、鹌鹑、麻雀、小鼠等8种动物的红细胞作血凝试验,结果证明,鸡传染性支气管炎病毒H_(120)株和M_(41)株均能凝集人O型以及羊、猪、兔、鸡、鸭、鹌鹑、小鼠的红细胞,而不能凝集麻雀的红细胞;GIBV株和Connecticut株能凝集人O型以及免、鸡、鹌鹑、麻雀的红细胞,而不能凝集猪、羊、鸭、小鼠的红细胞;Gray株能凝集兔、鸡、鹤鹑的红细胞,不能凝集人O型以及猪、羊、鸭、麻雀、小鼠的红细胞;而经家兔A型魏氏梭菌培养液处理的T株和未经处理的6株病毒对人O型以及8种动物的红细胞都没有凝集性。试验还证明,M_(41)株和H_(120)株对人O型及8种动物的血凝活性水平有差异。     

鸡传染性支气管炎病毒血凝抑制试验研究

用A型魏氏梭菌滤液处理鸡传染性支气管炎病毒(IBV)M41株,使其产生血凝性,成功建立IBV血凝抑制试验方法(IBV-HI).血凝效价为1:256～1:1024.抗原在2～8℃的保存期至少为5个月.用该抗原对IBV标准阳性血清、SPF鸡血清、其它鸡病标准阳性血清以及含M41株病毒的灭活苗免疫的鸡血清进行检测,证明IBV-HI试验具有很好的特异性.结果表明,该方法简便、快速、特异性强,可用于疫苗质量控制和疫苗免疫效力测定.     

鸡传染性支气管炎病毒血凝谱

用鸡传染性支气管炎病毒标准毒株Ｈ５２、Ｍ４１、Ｔ、Ｎ１１５、ＩＢＶ６２株及国内华中（ＨＺ）、华东（ＨＤ）、华南（ＨＮ）、华北（ＨＢ）、东北（ＤＢ）及西北（ＸＢ）等地方性分离株试验,研究ＩＢＶ血凝特性。结果表明,ＩＢＶ不直接凝集鸡、牛、兔、小白鼠、仓鼠、猪、鹅、马、绵羊及人的“Ｏ”型红细胞;１％胰蛋白酶３７℃作用ＩＢＶ４ｈ后,可凝集鸡、兔、小白鼠、仓鼠、鹅等动物红细胞;１０％乙醚３７℃作用ＩＢＶ２ｈ后,可凝集鸡、牛、小白鼠、仓鼠、鹅、绵羊及人的“Ｏ”型红细胞;１％卵磷脂酶Ｃ３７℃作用ＩＢＶ４ｈ后,可凝集鸡、兔、小白鼠、仓鼠、鹅等动物红细胞。     

不同免疫血清对鸡传染性支气管炎病毒血清型鉴定的影响

鸡传染性支气管炎病毒 ( avian infectious bron-chitis virus,IBV)由于其血清型众多 ,在预防该病时 ,常因所用疫苗株与流行株血清型不同而导致免疫失败[1,2 ] 。因而国内外学者普遍认为 ,IB研究的重点应放在 IBV血清型的鉴定和研制相应的弱毒疫苗和灭活疫苗上 [3 ]。国外已成功地将血凝抑制试验 ( HI)应用到不同 IBV毒株的血清分型上 ,King和 Hopkins[4 ] 证明HI试验是一种快速、简便的 IBV分型方法。但在应用该法进行 IBV分型时 ,常会遇到选择免疫血清的问题。本文比较了 3种免疫血清在 HI法中对 IBV血清分型的影响 ,得到了可…     

鸡传染性支气管炎病毒肾型毒株血凝特性的研究

本文对鸡传染性支气管炎病毒（ＩＢＶ）澳大利亚Ｔ株和分离的四株肾型毒株（ＢＪ＿（９３０１）、ＢＪ＿（９３０２）、ＴＪ＿（９３０１）、ＨＮ＿（９３０１）株）的血凝特性进行了系统研究。结果表明ＩＢＶ供试毒株经１～２％胰蛋白酶处理后，能凝集鸡和小鼠红细胞，同一毒株对两种红细胞的凝集价相近，且均能被ＩＢＶ澳大利亚Ｔ株血清所抑制。处理后的ＩＢＶ对鸡红细胞的血凝以缓冲液浓度０．０１～０．０５ＭｐＨ６．０～８．４为宜，对反应温度及稀释液种类无严格要求。ＩＢＶ的血凝活性能耐受３７℃２３天、５６℃１２小时、８０℃１．５小时，但在－３０℃、４℃和室温下保存２个月后，绝大部分丧失血凝活性。ＩＢＶ经０．２％甲醛、０．２％脱氧胆酸钠、２％硼氢化钠、０．０１Ｍ高碘酸钠３７℃灭活２４小时，仍能保持其血凝活性。从ＩＢＶ的血凝活性可被过量胰蛋白酶破坏和氯仿灭活，推断该血凝素可能是由脂质和蛋白质组成。     

鸡传染性支气管炎血凝抑制试验方法标准化的研究

鸡传染性支气管炎病毒（ＩＢＶ）抗体的血凝抑制（ＨＩ）试验方法经标准化后，检测ＳＰＦ鸡血清ＩＢＶ感染鸡血清以及野外样品的结果表明，具有快速，特异，敏感，稳定等优点。标准化的实现主要通过抗原冻干和鸡红细胞醛化，从而取得较为满意的结果。     

鸡传染性支气管炎病毒血凝抑制试验抗原的研制

为应用血凝抑制(HI)试验检验鸡传染性支气管炎疫苗免疫效力(血清、卵黄抗体水平),建立了鸡传染性支气管炎病毒HI试验抗原的制备方法。该方法是通过选取抗原谱最广的鸡传染性支气管炎病毒(IBV)M41株,经SPF鸡胚增殖培养36h后无菌收取鸡胚尿囊液,4℃、12 000r/min离心10min,上清用聚乙二醇(PEG)20000浓缩100倍;兔源A型产气荚膜梭菌中国标准株(C57-1株)37℃增殖培养18h,4℃、12 000r/min离心10min取上清,经PEG 20000透析袋浓缩5倍后通过0.20μm滤膜过滤除菌,然后将IBV液和A型产气荚膜梭菌菌液(含α毒素)二者按一定比例混合后经37℃恒温振荡感作2h,4℃经48h后制成。经大量试验表明,制备的IBV HI试验抗原效价高、稳定性好,可替代进口抗原应用于鸡群IBV疫苗免疫后血清抗体及卵黄抗体的HI效价检测。     

间接血凝试验检测鸡传染性支气管炎病毒抗体

间接血凝试验检测鸡传染性支气管炎病毒抗体江国托，徐宝春，王永坤（扬州大学农学院２２５００１）目前，鸡传染性支气管炎（ＩＢ）已成为国内养鸡业的主要疾病之一，虽然已建立的检测ＩＢＶ及其抗体的方法很多，如琼脂扩散试验、细胞中和试验、ＥＬＩＳＡ、血凝和血凝抑...     

鸡传染性支气管炎病毒(M41株)血凝抑制试验抗原的制备及检验

用传染性支气管炎病毒(IBV)M41株毒种接种11日龄鸡胚,收获感染鸡胚病毒液,将新鲜的病毒液用小型盒式超滤浓缩系统浓缩,然后用高速离心机离心,去上清,沉淀用原病毒液体积1/100的HA缓冲液悬浮,制备出100倍浓缩的IBV病毒液;将100倍浓缩的IBV病毒液中加入15%^20%的A型魏氏梭菌滤液,充分混匀,放37℃恒温振荡器感作2h,取出置2-8℃条件下48 h.制备出了5批IBV M41株HI杭原.检验结果显示:5批HI抗原的性状均为白色混浊液体,底部有少量沉淀;无菌检验为阴性,HA效价为1:128～1:512,5批杭原与IB阳性血清呈阳性反应,HI效价为81og2,而与阴性血清、新城疫阳性血清、H9亚型禽流感阳性血清、H5亚型禽流感阳性血清及产蛋下降综合征阳性血清呈阴性反应,HI效价低于21og2.各项检验结果均符合要求.     

Coronavirus avian infectious bronchitis virus

Infectious bronchitis virus (IBV), the coronavirus of the chicken (Gallus gallus), is one of the foremost causes of economic loss within the poultry industry, affecting the performance of both meat-type and egg-laying birds. The virus replicates not only in the epithelium of upper and lower respiratory tract tissues, but also in many tissues along the alimentary tract and elsewhere e.g. kidney, oviduct and testes. It can be detected in both respiratory and faecal material. There is increasing evidence that IBV can infect species of bird other than the chicken. Interestingly breeds of chicken vary with respect to the severity of infection with IBV, which may be related to the immune response. Probably the major reason for the high profile of IBV is the existence of a very large number of serotypes. Both live and inactivated IB vaccines are used extensively, the latter requiring priming by the former. Their effectiveness is diminished by poor cross-protection. The nature of the protective immune response to IBV is poorly understood. What is known is that the surface spike protein, indeed the amino-terminal S1 half, is sufficient to induce good protective immunity. There is increasing evidence that only a few amino acid differences amongst S proteins are sufficient to have a detrimental impact on cross-protection. Experimental vector IB vaccines and genetically manipulated IBVs--with heterologous spike protein genes--have produced promising results, including in the context of in ovo vaccination.     

鸡肾型传染性支气管炎病毒组织嗜性的研究

本研究用鸡肾型传染性支气管炎病毒 (肾型 IBV) C90 0 1株人工感染 14日龄 SPF鸡 ,于接种后定期采集病鸡的各组织器官制备成石蜡切片 ,用建立的检测石蜡切片中肾型 IB病毒的免疫酶组化染色技术 ,对人工感染肾型传染性支气管炎 (肾型 IB)鸡发病后病毒的组织器官亲嗜性和动态分布规律进行了研究。结果表明 ,肾型 IBV在胞浆内复制 ,主要亲嗜气管黏膜的上皮细胞和固有层腺体细胞、肺各级支气管上皮细胞以及肺房和呼吸性毛细管上皮细胞、气囊上皮细胞、肾和输尿管上皮细胞、消化道上皮和固有层腺体细胞、肝小叶间胆管上皮细胞、胰腺导管上皮细胞和腺泡上皮细胞、法氏囊淋巴滤泡髓质区淋巴细胞和网状细胞、胸腺小叶髓质区淋巴细胞和网状细胞、脾小体淋巴细胞、盲肠扁桃体弥散性淋巴组织的淋巴细胞以及心肌细胞 ,病毒出现的先后顺序为气管、肺脏、肾脏、输尿管 ,消化道、法氏囊、肝脏、胰脏、胸腺和气囊 ,盲肠扁桃体 ,脾脏 ,心肌。病毒在肾脏和输尿管持续 2 0天 ,气管为 13天 ,消化道和法氏囊为 11天 ,肝脏、胰脏和肺为 10天 ,胸腺为 6天 ,气囊为 5天 ,盲肠扁桃体、脾脏、心肌呈一过性感染     

Recent studies on the enterotropic strain of avian infectious bronchitis virus

Avian infectious bronchitis (AIB) is an economically important disease of chickens. Recent studies have revealed enterotropism by at least one strain of AIB virus with pathological lesions in parts of the gut. This review highlights the findings of the studies so far made on this enterotropic strain of AIB virus.     

Chicken embryonal vaccination with avian infectious bronchitis virus

A commercial infectious bronchitis virus (IBV) vaccine of the Massachusetts 41 strain was injected in embryonating chicken eggs on embryonation day (ED) 18. The IBV vaccine was pathogenic for embryos, and it was passaged in chicken kidney tissue culture to reduce the pathogenicity. At the 40th tissue culture passage (P40-IBV), the virus became apathogenic for the embryos. Maternal antibody-positive or -negative chicks hatching from eggs injected with P40-IBV developed antibody to IBV and were protected against challenge exposure at 4 weeks of age with virulent Massachusetts 41 IBV. Although P40-IBV protected chicks when administered on ED 18, this virus did not protect chicks well if given at hatch. When combined with the turkey herpesvirus (HVT), P40-IBV given on ED 18 did not interfere with the protection against challenge exposure with virulent Marek's disease virus, nor did the presence of HVT interfere with protection by P40-IBV. Thus, under laboratory conditions, IBV vaccine could be combined with HVT to form a bivalent embryonal vaccine.     

1株QX基因型鸡传染性支气管炎病毒的致病性

通过对8日龄SPF鸡人工感染鸡传染性支气管炎病毒(IBV)QX株发病,分别在攻毒后3,7,30 d剖杀并采集气管、肺、肾、脾、胸腺、法氏囊制作病理组织切片。攻毒7 d后,采集上述器官相同部位进行免疫组织化学法分析。根据IBV-QX株NP基因的保守序列设计引物,构建质粒标准品,建立了IBV-QX Real-time PCR标准曲线,并跟踪检测试验鸡的口腔、泄殖腔排毒情况。此外,观察并记录了2组鸡的临床症状、体质量差异等信息。结果显示,发病初期(3 d),气管、肺几乎无明显病理变化,其他器官均出现不同程度的炎性反应;临床症状明显期(7 d),所涉及的器官均有不同程度的病理变化;发病后期(30 d),气管纤毛有所恢复,肺、脾炎性反应有所减轻,但肾脏和法氏囊的病变依然严重。免疫组织化学法检测到上述器官均有可见抗原聚集。30 d时被感染鸡的口腔、泄殖腔仍可检出病毒核酸。试验鸡临床症状明显,体质量差异显著。本试验旨在探究具有组织嗜性的IBV-QX毒株对宿主的免疫器官和该病毒的主要靶器官的病理损伤以及被损伤器官的自我修复情况,抗原分布差异,排毒规律等,为后续研究宿主免疫器官发挥天然免疫的相关机理打下基础,也为研发新的疫苗、相关药物的临床药效评价、制定合理的疫苗接种策略提供参考。     

Attenuation of avian infectious bronchitis virus by cold-adaptation.

Avian infectious bronchitis virus (IBV) Arkansas-type DPI strain was passaged 10 times in specific-pathogen-free (SPF) chicken embryos incubated at 28 C and 37 C. Virus grown at 28 C acquired cold-adapted (CA) and temperature-sensitive (TS) characteristics based on more-rapid growth at 28 C and a reduced ability to grown at 41 C, respectively, compared with non-cold-adapted (non-CA) virus grown at 37 C. The pathogenicity and immunogenicity were determined for CA and non-CA IBV in 1-day-old SPF chickens following intratracheal inoculation. The percentage of CA IBV-vaccinated chicks exhibiting respiratory disease exceeded 30% on only 1 day postinoculation (PI) (day 5 PI), compared with 8 days (days 2-9 PI) for birds given non-CA IBV. Mortality was 0% for CA IBV-vaccinated chickens and 6% for non-CA virus-vaccinated chickens. Microscopically, both CA and non-CA IBV caused diffuse tracheal deciliation, although mucosal hyperplasia, necrosis, and heterophil infiltration were more severe with non-CA IBV. Virus was reisolated from kidneys of chickens given CA IBV, suggesting the loss of the TS property. The instability of the TS property was confirmed by growth of the reisolated virus at 41 C. Both CA and non-CA viruses induced complete protection against homologous challenge virus infection of the upper respiratory tract.