共查询到19条相似文献，搜索用时 218 毫秒
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《中国家禽》2016,(16)
为比较Vero、DF-1和MDCK三种传代细胞对鸡传染性法氏囊病病毒(IBDV)BJQ902株的敏感性。研究采用盲传的方法,将BJQ902株分别接种Vero、DF-1和MDCK细胞,连续传代至第8代(F8),收获每代次的病毒液测定TCID50,同时应用SYBR GreenⅠ实时荧光定量RT-PCR法检测病毒拷贝数,分析不同细胞对BJQ902株的敏感性。结果表明经过8代盲传后,收获的IBDV BJQ902株在DF-1细胞、Vero细胞和MDCK细胞培养的病毒液,使用CEF细胞测定病毒滴度分别为107.9TCID50/0.1 m L、106.5TCID50/0.1 m L和103.7TCID50/0.1 m L;SYBR GreenⅠ实时荧光定量RT-PCR法检测表明IBDV在DF-1适应传代6代后,培养液的病毒拷贝数在106.5μL以上,而IBDV抗原拷贝数在Vero细胞和MDCK细胞培养液的拷贝数各代次均低于DF-1细胞。由此可见DF-1细胞对IBDV BJQ902株的敏感性高于Vero细胞和MDCK细胞。 相似文献
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试验比较了MA-104、Marc-145、Vero 3种猴肾细胞增殖犬瘟热病毒的敏感性,按常规方法将犬瘟热病毒(CDV-XN112株)分别接种于MA-104、Marc-145、Vero 3种细胞,通过观察细胞病变,确定收毒时间,比较病毒滴度。结果表明,犬瘟热病毒可在MA-104、Marc-145、Vero 3种细胞内增殖,并出现典型细胞病变,效价均达到104.5TCID50/0.1mL以上,而Vero细胞增殖犬瘟热病毒更为经济高效,是增殖该病毒的理想细胞。 相似文献
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本研究以伪狂犬病病毒(pseudorabies virus,PRV)Ra株体外感染不同的细胞系为模型,研究PRV Ra株最适细胞系。试验采用ST细胞、PK-15细胞、Vero细胞、F81细胞、DF1细胞、MDCK细胞和CEF细胞7种细胞作为该毒株的接种对象,观察毒株在这几种细胞上病变特点、细胞病变时间及病毒增殖规律等并进行比较。观察结果发现,受试的各种细胞在PRV感染期间均能表现明显的细胞病变,但不同细胞在病变时间上存在较大差异,其中PK-15和ST细胞在感染后24 h内出现明显细胞病变,时间最早。TCID50测定病毒增殖力结果表明,ST细胞在病毒增殖传代中毒力保持最好,与原毒株毒力相当,为106.9 TCID50/0.1 mL。本试验结果表明,ST细胞是较适合于PRV Ra株体外研究的细胞系。 相似文献
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犬瘟热病毒的分离及部分特性研究 总被引：2，自引：0，他引：2
本试验通过消除组织培养中某些干扰因素,用鸡胚成纤维细胞(CEF)直接从水貂病料中分离出3株犬瘟热病毒,即QM—CDV、YM—CDV和ZM—CDV,并以3株国外犬瘟热弱毒作为参考株,对所分离毒株的部分生物学特性进行了研究.结果表明,QM—CDV、YM—CDV和ZM—CDV在鸡胚成纤维细胞上生长良好,产生明显的细胞病变(CPE),也可在猴肾传代细胞(Vero)和人羊膜细胞(FL)上增殖并出现细胞病变;在电镜下观察,病毒粒子具有囊膜和纤突,直径在130～180nm之间;细胞中和试验表明,分离毒与国外弱毒具有相同的血清型;分离毒株的水貂病料经脑内接种均使幼犬发病,出现明显的临床症状. 相似文献
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《中国兽医学报》2015,(7):1046-1050
以犬瘟热病毒(CDV)的核衣壳蛋白基因(N基因)和血凝素蛋白基因(H基因)作为靶基因,各设计并体外合成小干扰RNA,分别为N1、N2、N3和H1、H2、H3。将6对siRNA分别转染非洲绿猴肾细胞(Vero)并在转染后接种CDV,48h后通过细胞病变(CPE)、间接免疫荧光、TCID50、荧光定量PCR等方法对2组6个siRNA抑制CDV在Vero细胞中增殖的效果进行比较研究。结果显示,转染细胞感染CDV 48h后,6个siRNAs干扰组细胞出现的绿色荧光明显少于对照组,病毒感染滴度分别是对照组的1/293、1/11、1/29、1/83、1/302和1/90,6个干扰组的CDV基因拷贝数分别是对照组的12.96%、57.75%、23.47%、31.82%、12.82%、15.58%。结果表明,6对siRNAs均对CDV在Vero细胞中的复制具有不同程度的抑制作用,且以H2组抑制效率最高。 相似文献
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《畜牧与兽医》2015,(8):89-92
为了探索猪流行性腹泻病毒(PEDV)在微载体培养Vero细胞上的增殖效果,采用2 L生物反应器进行微载体培养Vero细胞和PEDV繁殖,并摸索细胞生长和病毒繁殖的最佳条件,检测病毒滴度,与方瓶培养的病毒滴度进行比较。结果显示:当微载体为5 g,种子细胞数约3.05×107个,采用灌注式培养,Vero细胞经72 h长满单层,细胞数约为3.02×109个,此时用滴度为104.45TCID50/m L的PEDV 1 m L感染Vero细胞,培养96 h,微载体上80%细胞脱落时收毒,培养的PEDV具有较高的病毒滴度。相同病毒在微载体系统与方瓶上进行同步传代,经检测病毒滴度均有提高,但在微载体系统上培养的PEDV最高滴度可达到106.05TCID50/m L,明显高于方瓶培养。本试验为研究PEDV能更好适应Vero细胞,提高PEDV产量提供技术支持。 相似文献
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为了研究绿猴肾(Vero)细胞中禽呼肠孤病毒(ARV)的增殖特性,试验采用了光学显微镜观察、半数组织感染量(TCID50)测定、RT-PCR检测等方法对细胞培养物进行了研究。结果表明:ARV S1133毒株在Vero细胞中能有效增殖,并出现细胞破碎、萎缩、抱团、脱落、边缘模糊、胞体变大,甚至死亡等典型的细胞病变(CPE);RT-PCR检测成功扩增出一条大小为1 248 bp的条带;ARV在感染Vero细胞后会经历三个时期,即潜伏期、快速增长期、稳定期,并在稳定期病毒效价达到峰值,TCID50为1×10~(-8.46)/0.1 m L。 相似文献
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将猪细小病毒(PPV)接种于F81和PK15细胞,研究不同的培养基、pH值、血清含量、细胞密度对病毒在细胞里增殖的影响,细胞毒液冻融3次后测半数组织细胞感染量(TCID50)。试验结果表明,接毒24 h后F81和PK15细胞都有病变产生,在细胞病变达到75%以上时收获细胞毒液,发现PPV在F81细胞和PK15细胞中均能增殖且病毒滴度比较高。测得F81细胞接种PPV最高滴度为108.71TCID50/mL;PK15细胞接种PPV最高滴度为108.73TCID50/mL。 相似文献
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为探讨新城疫Ⅳ系和Ⅰ系毒株对鸡胚成纤维细胞(CEF)致病变特性的影响,采用细胞病变观察(CPE)、血凝试验(HA)和半数细胞培养物感染量(TCID50)测定等方法对新城疫Ⅳ系和Ⅰ系毒株在CEF上的感染特性进行检测。结果表明:经鸡胚复壮的新城疫Ⅰ系毒株不管胰蛋白酶是否参与,均能在CEF上增殖并产生典型的细胞病变,主要以发生细胞融合和形成多核巨细胞为特征;而新城疫Ⅳ系毒株,必须在胰蛋白酶参与的情况下才能产生CPE,主要以胞浆内出现可见大小不等的空泡或颗粒为特征。新城疫Ⅰ系毒株与新城疫Ⅳ系毒株(胰酶参与)相比CEF培养上清液最高HA效价高2个滴度,TCID50也明显增高,证明新城疫Ⅰ系毒株在CEF细胞上的感染力更强。 相似文献
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Lan NT Yamaguchi R Kai K Uchida K Kato A Tateyama S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(5):491-495
To know growth profiles of canine distemper virus (CDV) on Vero cells stably expressing canine signaling lymphocyte activation molecule (Vero-DogSLAMtag; Vero-DST cells), the propagation of three strains of CDV was tested in Vero-DST cells in comparison with parental Vero cells. Strain MD77 could grow well in both cell lines, but demonstrated no syncytium formation or indistinguishable rounding cytopathic effects (CPE) in Vero cells. Strains Onderstepoort and KDK-1 also grew well in Vero-DST cells with apparent syncytium CPE, while they grew less or no efficiently, respectively, in Vero cells. All three CDV strains demonstrated the peak titers, in Vero-DST cells before reaching to an extensive CPE and drastic decrease of titers at/after full CPE. Immunohistochemistry revealed that viral antigens of all CDV strains were found exclusively in the syncytia in Vero-DST cells, while in Vero cells, viral antigen was identified in their single cells for strain MD77 but none for other strains. Thus, every strain of CDV could grow well in Vero-DST cells and behaved differently against Vero cells. These results would be of practical value for workers of CDV because 1) In Vero-DST cells, by observation of distinct syncytium CPE, the highest titer or the best growth of virus could be identified; 2) In Vero cells, various CDV strains could be readily classified after propagation in Vero-DST cells. 相似文献
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取山东某貂场疑似犬瘟热病毒（canine distemper virus,CDV）感染的水貂肝脏等组织病料,通过RT-PCR检测呈CDV阳性,且无水貂细小病毒存在,将病料接种原代CEF细胞、传代系Vero细胞和DF1细胞3种细胞进行病毒分离,通过优化细胞培养条件,最终在Vero细胞上传代培养成功,出现露珠状典型细胞病变（CPE）。分离毒应用RT-PCR、PCR产物测序、抗体中和试验（SN）、间接免疫荧光试验（IFA）及病毒包涵体检查等多种方法进行了CDV的鉴定,结果显示CDV阳性,表明分离到的病毒为水貂CDV,并将其命名为CDV LD-1株。 相似文献
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Guo A Lu C 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2000,47(3):183-190
Apoptosis of Vero cells infected with two canine distemper virus (CDV) vaccine strains was detected using TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick end-labelling (TUNEL), flow cytometric analysis, agarose gel electrophoresis and electron microscopy (EM). By TUNEL, apoptotic cells were found in CDV-Onderstepoort (CDV-Ond)-infected cells. DNA fragments isolated from infected cells were separated by agarose gel electrophoresis and a 'ladder' pattern appeared. EM observations demonstrated that the cells undergoing cytopathic effect (CPE) possessed morphological characteristics of apoptotic cells. Flow cytometric analysis indicated that CDV could induce apoptosis of Vero cells, but the percentages of the apoptotic cells were correlated with the CPE types. The strain showing the cell-rounding type of CPE produced a much higher percentage of apoptotic cells than CDV-Ond with the syncytium type of CPE (P < 0.01). It was concluded that CDV vaccine strains could induce apoptosis of Vero cells and the apoptosis was virus strain-dependent and cell-dependent. The mechanism remains to be studied. 相似文献
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对流行病学调查、临床症状检查和ELISA检测为犬瘟热阳性的自然发病犬,取肠内容物为病料,采用同步培养方法接种于犬肾细胞系(MDCK)进行病毒的分离,并对分离株进行了形态学特征、血凝特性、动物感染及RT-PCR鉴定。结果表明:病料接种MDCK细胞产生明显的细胞病变(CPE),电镜负染观察接毒细胞培养物见有典型的犬瘟热病毒粒子。分离株不凝集鸡及人“O”型红细胞,接种犬出现明显的临床症状和病理变化。用RT-PCR技术检测病毒细胞培养液,扩增出的片段长为760 bp,与预期设计的长度相同,由此确证分离株为犬瘟热病毒,命名为CDV-GZ2株。 相似文献
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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. 相似文献
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Growth of serotypes I and II and variant strains of infectious bursal disease virus in Vero cells 总被引：4，自引：0，他引：4
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. 相似文献
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Serageldeen Sultan Nguyen Thi Lan Toshiki Ueda Ryoji Yamaguchi Ken Maeda Kazushige Kai 《Acta veterinaria Scandinavica》2009,51(1):38
Backgrounds
The aim of this study was to confirm the propagation of various canine distemper viruses (CDV) in hamster cell lines of HmLu and BHK, since only a little is known about the possibility of propagation of CDV in rodent cells irrespective of their epidemiological importance.Methods
The growth of CDV in hamster cell lines was monitored by titration using Vero.dogSLAMtag (Vero-DST) cells that had been proven to be susceptible to almost all field isolates of CDV, with the preparations of cell-free and cell-associated virus from the cultures infected with recent Asian isolates of CDV (13 strains) and by observing the development of cytopathic effect (CPE) in infected cultures of hamster cell lines.Results
Eleven of 13 strains grew in HmLu cells, and 12 of 13 strains grew in BHK cells with apparent CPE of cell fusion in the late stage of infection. Two strains and a strain of Asia 1 group could not grow in HmLu cells and BHK cells, respectively.Conclusion
The present study demonstrates at the first time that hamster cell lines can propagate the majority of Asian field isolates of CDV. The usage of two hamster cell lines suggested to be useful to characterize the field isolates biologically. 相似文献19.
为获得大熊猫犬瘟热病毒株,采集死亡大熊猫的心脏、肺、肝病料,研磨并反复冻融后收集上清液接种Vero细胞,待出现细胞病变(CPE)后,收集病毒液。用逆转录-聚合酶链反应(RT-PCR)鉴定病毒分离株,测定病毒TCID_(50),动物回归试验测定其毒力。结果显示,盲传到第5代时,Vero细胞出现圆缩、聚集、脱落等病变;PCR扩增出287bp片段,与预期相符;测序结果显示,该毒株与已发表的CDV SD(14)11毒株(亚洲-Ⅰ型)的同源性为98%;毒株的TCID_(50)为10~(-5.2)/mL;幼犬感染分离毒株后出现体温升高、鼻头发干、拉稀、眼鼻分泌出水样分泌物等症状。本研究成功分离出大熊猫源犬瘟热病毒株。 相似文献