鸡传染性法氏囊病病毒适应于Vero细胞的初步研究

将国内６个省市（广东、巾东、河北、辽宁、、新疆和北京）的７株鸡传染性法氏囊病（ＩＢＤ）的法氏囊组织处理后，直接在Ｖｅｒｏ细胞上盲传３～４代，均能不同程度的产生特征性细胞病变效应（ＣＰＥ），并通过选用具有广谱性的抗法氏囊病病毒的单克隆抗体（ＩＢＩ）Ｖ－ＭｃＡｂ）采用间接免疫荧光（ＩＦＡ）和碱性磷酸酶—抗碱性磷酸酶桥联酶显色技术（ＡＰＡＡＰ）的方法对７株分离毒的Ｖｅｒｏ第８代细胞进行鉴定，又用逆转—聚合酶链式反应（ＲＴ－ＰＣＲ）对７株分离毒的第１４代Ｖｅｒｏ细胞毒进行进一步证实，结果表明这７株ＩＢＤ囊毒均适应于Ｖｅｒｏ细胞上，这是国内首次报道将组织毒直接适应于Ｖｅｒｏ传代细胞上。     

适应Vero细胞传染性腔上囊病毒X毒株对鸡胚的致病性试验

适应Ｖｅｒｏ细胞传染性腔上囊病毒（ＩＢＤＶ）Ｘ毒株,通过对鸡胚的致死作用、病理变化的结果表明,ＩＢＤＶ Ｘ毒株在Ｖｅｒｏ细胞上传代后,各试验代次毒株均能适应鸡胚,并引起鸡胚死亡,对鸡胚的致病性随传代次数的增加而减弱。     

新城疫病毒在不同的细胞上增殖特性的研究

研究新城疫病毒（Ｎｅｗｓｃａｓｔｌｅ ｄｉｓｅａｓｅ Ｖｉｒｕｓ,ＮＤＶ）可,弱毒株在不同的传代细胞的生物学特性。Ｆ４８Ｅ８强毒株在ＢＨＫ、Ｖｅｒｏ和ＣＥＦ中都能增殖并产生细胞病变,而Ｖ４生弱毒株不能在ＢＨＫ和Ｖｅｒｏ中增殖,当ＭＥＭ含１０ｕｇ／ｍＬ胰蛋白酶时,ＮＤＶ弱毒株在Ｖｅｒｏ中能较好地复制,将Ｖ４株在Ｖｅｒｏ中传代培养后,Ｖｅｒｏ中传代的Ｖ４毒株的ＨＡ较低,但毒力增强,ＢＨＫ中传代的Ｖ４毒     

CDV93039株在Vero细胞上增殖的研究

ＣＤＶ９３０３９株感染Ｖｅｒｏ细胞后主要表现细胞变性、坏死，空泡化、合胞体形成和包涵体的出现。感染后４天可见胞浆包涵体，８天可见核内包涵体，核内包涵体的产生可能与病毒的毒力有关。与文献报道的ＣＤＶ其他毒株相比，ＣＤＶ９３０３９株在Ｖｅｒｏ细胞上的感染速率属中间型。ＣＤＶ９３０３９株在Ｖｅｒｏ细胞上增殖后的毒力较高，除不同毒株间可能存在差异外，培养温度的高低亦是原因之一。ＣＤＶ９３０３９株在Ｖｅｒｏ细胞上的最佳增殖条件是在３３℃培养８～１２天。     

鸡传染性支气管炎病毒(IBV)干扰新城疫病毒(NDV)增殖现象的研究

ＩＢＶ毒株Ｈ１２０、Ｈ５２、ＭＡ５对ＮＤＶ－ＬａＳｏｔａ的干扰实验证明ＩＢＶ特异性干扰ＮＤＶ增殖。１）采取不同顺序的同胚接种法：ＩＢＶ接种之后再接种ＮＤＶ;或ＮＤＶ接种之后再接种ＩＢＶ,及ＩＢＶ,ＮＤＶ同时接种,ＩＢＶ均干扰ＮＤＶ的增殖。２）ＮＤＶ接种３６小时之内,干扰现象最为明显,Ｈ１２０,Ｈ５２的干扰能力稍强于ＭＡ５。３）ＮＤＶ血清中和实验结果显示,不同顺序同胚接种ＮＤＶ、ＩＢＶ时,ＮＤＶ－ＬａＳｏｔａ不影响ＩＢＶ的增殖能力。同胚增殖ＩＢＶ,ＮＤＶ的关键是控制ＮＤＶ、ＩＢＶ的接毒量及选择合适的收毒时间。     

鸡传染性法氏囊病Vero细胞弱毒疫苗（VCV－901）兔疫鸡的抗体检测

鸡传染性法氏囊病Ｖｅｒｏ细胞弱毒疫苗（ＶＣＶ－９０１）兔疫鸡的抗体检测马兴树（河北邯郸农专牧医系）近年来，国内外养鸡生产者对鸡传染性囊病（ＩＢＤ）的防制工作极为重视，我国对ＩＢＤ的疫苗研究也取得了一定进展，但ＩＢＤ弱毒疫苗的生产均由鸡胚或鸡胚细胞制备...     

应用鸡胚法氏囊原代细胞培养鸡IBDV的研究

试用鸡胚法氏囊原代细胞传代、增殖鸡传染性法氏囊病病毒（ＩＢＤＶ），均不同程度地产生特征性细胞病变效应。采用双抗体夹心法ＥＬＩＳＡ和细胞毒回归鸡试验证实，ＩＢＤ－ＶＨ株组织毒适应于鸡胚法氏囊细胞，并随传代次数增加，病毒增殖能力增强；而在鸡胚成纤维细胞上盲传２代之后才开始出现细胞病变效应。比较了ＩＢＤＶ在鸡胚法氏囊细胞和鸡胚成纤维细胞上增殖能力的差异，讨论了培养基ＰＨ值、小牛血清浓度对鸡胚法氏囊细胞培养及病毒增殖的影响。     

CDV93039株在Vero细胞上增殖的研究

ＣＤＶ９３０３９株感染Ｖｅｒｏ细胞后主要表现细胞变性，坏死，空泡化，合肥体形成和包涵体的出现。感染后４天可见胞浆包涵体，８天可见核内包涵体，核内包涵体的产生可能与病毒的毒力有关。与文献报道的ＣＤＶ其他毒株相比，ＣＤＶ９３０３９株在Ｖｅｒｏ细胞上的感染速率属中间型。     

FMDV在BHK21细胞扩增后FMDV 140 S抗原的测定

本文报告了用蔗糖密度梯度离心和光密度分析相结合口蹄疫病毒（ＦＭＤＶ）在单层ＢＨＫ２１细胞中扩增后，其中ＦＭＤＶ １４０ Ｓ抗原浓度，并同时分析了经ＢＥＩ（二乙烯亚胺）灭活后，这种抗原含量的改变。结果表明，Ｏ型ＦＭＤＶ在单层ＢＨＫ２１细胞中增殖后，其ＦＭＤＶ １４０ Ｓ抗原浓度降至０．９μｇ／ｍｌ。     

应用鸡胚法氏囊原代细胞培养鸡IBDV的研究

试用鸡胚法氏囊奈代细胞传代，增殖鸡传染性法氏囊病病毒（ＩＢＤＶ），均不同程度地产生特征性细胞病变效应，采用双抗体夹心法ＥＬＩＳＡ和细胞毒回归鸡试验证实，ＩＢＤＶＨ株组织感毒适应于鸡胚法氏囊细胞，并随传代次数增加，病毒增殖能力增强，而在鸡胚成纤维细胞上盲传２代之后才开始出现细胞病变效应，比较了ＩＢＤＶ在鸡胚法氏囊细胞和鸡胚成纤维细胞上增殖能力的差异，讨论了培养基ＰＨ值，小牛血清浓度对鸡胚法氏囊细胞培     

Replication of infectious bursal disease virus in continuous cell lines

Three mammalian continuous cell lines--MA-104, Vero, and BGM-70--were tested for their ability to support replication of infectious bursal disease virus (IBDV). Selected tissue-culture-adapted vaccine strains and tissue-culture-adapted field isolates of IBDV replicated in the MA-104, Vero, and BGM-70 cells; cytopathic effects were most pronounced in the BGM-70 cells. The cytopathic effects of the viruses in BGM-70 cells and chicken embryo fibroblast (CEF) cultures were similar. Virus-neutralization titers of selected serum samples determined in BGM-70 cultures compared well with those obtained from CEF cultures.     

Isolation and propagation of infectious bursal disease virus using the ovine kidney continuous cell line.

Twenty-six samples known to contain infectious bursal disease virus (IBDV) were examined by virus-isolation attempts on ovine kidney (OK) cell line, Vero cell line, and chicken embryo fibroblast (CEF) cultures. Virus was isolated from two of 26 samples, three of 26 samples, and three of 25 samples on OK, Vero, and CEF cultures, respectively. However, in contrast to IBDV replication in Vero and CEF cultures, isolated virus was unable to induce serially sustained cytopathic effects (CPE) during successive passages in the OK cell line, unless cell lysates were treated with chloroform between every other passage. The cytopathogenicity of the untreated virus passaged in OK cells was revived and maintained upon passage in Vero cells. An initial single passage of laboratory or field material in OK cells followed by further passages in Vero cells resulted in virus isolation from six of 26 samples, which was better virus recovery than when either cell line was used alone or when CEF cultures were used. Twenty of the 26 test samples were originally positive when examined by nucleic acid hybridization with radiolabeled IBDV cDNA, indicating that some of the samples that were negative upon virus isolation using OK and Vero cells may have contained inactivated virus.     

In situ localization of infectious bursal disease virus-binding cells by a biotin-streptavidin system.

A biotin-streptavidin system was established to directly visualize infectious bursal disease virus (IBDV)-binding cells in cell culture or in fresh tissues. The cells or tissue sections were first incubated with a biotinylated, purified IBDV strain GZ911 and then with a streptavidin-beta-galactosidase conjugate. In the presence of the enzyme substrate X-gal, IBDV-binding cells were labeled in blue color. By applying this method to frozen tissue sections, virus-binding sites were localized in situ in the bursa, spleen, and kidney tissue sections, whereas no positive cells were detected in the thymus tissue sections. Chicken embryo fibroblasts, Vero cells, MOP-8 cells, 293-EBNA cells, PANC-1 cells, and HuTu 80 cells were found to bind to the virus. However, the binding of the virus to MDA-MB-231 cells and SVG p12 cells was undetectable. This method can be employed for the expressional cloning of IBDV receptor and can be applied to studies on other avian viruses.     

Growth of serotypes I and II and variant strains of infectious bursal disease virus in Vero cells

The growth of five strains of infectious bursal disease virus--three strains of serotype I (SAL, D-78, 2512), one of serotype II (OH), and one variant strain (Variant-A)--were compared in Vero and chicken embryo fibroblast (CEF) cell cultures in order to characterize the replication of different strains of IBDV in Vero cells. For all five virus strains, the latent period in Vero cells ranged from 12 to 18 hr, which was longer than the 4-to-6-hr latent period observed in CEF cultures for strains SAL, D-78, and OH. Virus strains SAL, D-78, and OH, which were examined in both Vero and CEF cultures, also had a more extensive maturation phase and higher yields of virus in Vero than in CEF cultures. Total titers of these viruses of 5.35 to 6.10 log10 TCID50/ml in CEFs occurred 24 to 30 hr postinoculation (PI), although the cytopathic effect (CPE) was not seen until 72 hr PI. By comparison, their total infectious virus titers of 6.85 to 8.35 log10 TCID50/ml in Vero cells occurred from 48 hr PI, coinciding with the appearance of CPE. The growth curve of Variant-A in Vero cells differed from the other viruses by showing steadily rising extracellular and cell-associated virus titers throughout the 72-hr observation period. Only very low titers of Variant-A were obtained in CEF cultures, and thus no growth curve in CEFs was performed.     

Isolation of infectious bursal disease virus from turkeys with arthritic and respiratory symptoms in commercial farms in Quebec.

Infectious bursal disease viruses (IBDVs) were isolated from turkeys showing symptoms of arthritis and respiratory disease in commercial poultry farms in the province of Quebec, Canada. Synovial fluids collected from hock joints of arthritic birds and peripheral blood leukocytes obtained from the birds with respiratory problems were used for virus isolation in embryonated chicken eggs, and Vero and BGM-70 cell cultures. The infected cells were evaluated for the presence of IBDV by indirect immunofluorescence assay using monoclonal antibodies. The viruses were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of viral genome and by electron microscopy. Although one of these turkey isolates tested was neutralized by serotype 1-specific commercial chicken antisera, preliminary results indicated that there are antigenic differences between the Quebec isolate, IBDV QT-1, and the existing strains of IBDV belonging to serotype 1.     

Selection of an infectious bursal disease virus mutant with increased immunogenicity following passage under humoral immune pressure.

It is generally known that the pathogenicity of infectious bursal disease virus (IBDV) strains decreases following passage in cell culture. However, there is no information about the effect of passage under immune pressure on the phenotypic and molecular properties of IBDV. In the present study, a small plaque mutant virus with poor neutralization capability, but showing similar growth characteristics as the parental virus strain, QC2, was isolated after serial passage in Vero cells in presence of IBDV serotype 1 chicken polyclonal antiserum. This mutant virus showed reduced pathogenicity but enhanced immunogenicity compared to the parental virus. Sequence analysis of the non-coding regions of the genome revealed 4 and 3 nucleotide changes in the 3' non-coding regions of segments A and B, respectively, and none in the 5' non-coding regions. Restriction enzyme analysis of selected coding regions of the IBDV genome in both viruses revealed a loss of the PstI site in the VP2 region of the mutant virus. Selection of such mutant viruses by passaging under immune pressure may offer an improved method for developing safer and more effective attenuated vaccine strains against infectious bursal disease of chickens.     

Study on Proliferation Technology of Chicken Infectious Bursal Disease Virus BJQ902 Strain in DF-1 Cell Line

In order to produce high titers of infectious bursal disease virus(IBDV) antigen,the proliferation technology of chicken IBDV BJQ902 strain in DF-1 cell line was studied by the selection and optimization of the following five culture conditions,including the amount of inoculated virus,harvest time,inoculation time,culture temperature and serum concentration in maintenance media.The results showed that the optimal proliferation conditions of IBDV BJQ902 strain in DF-1 cell line were obtained as follow:The culture temperature was 37℃,the inoculum concentration was between 0.01% and 0.1% (V/V),the inoculation time was between 48 and 72 h after cell passage,the serum concentration in maintenance media was between 1% and 2%,the harvest time was between 60 and 72 h after inoculation.Under the optimal conditions,the virus titers were between 10^8.3 and 10^8.7 TCID₅₀/0.1 mL.     

鸡传染性法氏囊病病毒BJQ902株在DF-1细胞系上增殖工艺研究

试验旨在通过DF-1细胞增殖获得高效价的鸡传染性法氏囊病病毒（infectious bursal disease virus,IBDV）抗原。通过病毒接种量、收毒时间、接毒时间、温度和维持液血清浓度5个培养条件的筛选和优化,对IBDV BJQ902株在DF-1细胞上的增殖工艺进行了研究。结果表明,IBDV BJQ902株在DF-1细胞上最佳增殖条件为：在37℃条件下培养,接毒量在0.01%~0.1%之间,接种时间为细胞传代后生长48~72 h,维持液血清浓度为1%~2%,收毒时间为病毒接种后60~72 h。在此培养条件下增殖病毒毒价在10^8.3~10^8.7 TCID₅₀/0.1 mL之间。     

The Characterization of Infectious Bursal Disease Virus Strains/Isolates from Field Outbreaks in India

Pahar, B. and Rai, A., 1997. The characterization of infectious bursal disease virus strains/isolates from field outbreaks in India. Veterinary Research Communications, 21 (4), 289-301Three infectious bursal disease virus (IBDV) isolates were adapted to culture in chick embryo fibroblast cells in which they produced a cytopathic effect. The isolates were identified as IBDV by virus neutralization tests using a standard hyperimmune serum against infectious bursal disease, physicochemical properties and their pathogenicity in chick embryos and chicks. The IBDV S394 strain was antigenically different from IBDV S194/IBDV S494 as well as from the IBDV Intermediate Georgia strain, one of the vaccine strains in use in India.