人参皂苷及其衍生物抗马立克氏病毒的作用

为探讨人参皂苷及其衍生物体外抗马立克氏病毒(MDV)的作用机理,采用空斑试验测定人参皂苷及其衍生物对马立克氏病毒感染鸡胚成纤维细胞(CEF)的保护效果,结果发现人参皂苷及其衍生物均具有较好的抗病毒效果,特别是人参皂苷衍生物组增殖抑制的抗MDV空斑减少率达到盐酸吗啉胍的77%(55.62/72.52)。表明人参皂苷及其衍生物可以减轻病毒对CEF细胞的损伤程度。     

人参皂苷及其衍生物体内抗马立克氏病毒的间接免疫荧光观察与PCR检测

为了探讨人参皂苷及其衍生物体内抗马立克氏病毒的作用机理.用马立克氏病毒人工感染雏鸡模型,采取人参皂苷及其衍生物口服,通过间接免疫荧光试验动态观察人参皂苷及其衍生物能否减少MDV抗原在组织中的分布;通过PCR检测看人参皂苷及其衍生物能否减少MDV的出现.结果显示:人参皂苷组、衍生物组被检组织的阳性细胞数量明显比盐酸吗啉胍阳性对照组被检组织的阳性细胞数量少;MDV病毒核酸的PCR检测显示,药物没有阻止病毒对组织的感染.结果表明,人参皂苷及其衍生物抗马立克氏病毒效果要优于盐酸吗啉胍.     

分子修饰前后人参皂苷体外抗马立克氏病毒鸡胚成纤维细胞模型的建立

用雏鸡将马立克氏病毒(MDV)血毒复壮,分离发病鸡淋巴细胞并接种于鸡胚成纤维细胞,观察其病变。获得适应鸡胚成纤维细胞(CEF)的MD强毒,通过电镜观察、琼脂扩散实验进一步鉴定病毒,并建立了MDV感染CEF细胞模型,为研究人参皂苷及其衍生物的体外抗病毒作用及其机制奠定基础。     

人参皂苷及其衍生物抗马立克病病毒作用的超微结构和病理变化观察

为了探讨人参皂苷及其衍生物抗马立克氏病病毒的作用机制.采用马立克氏病毒感染的雏鸡作为模型,人参皂苷及其衍生物口服给药途径,动态观察不同时间段,试验鸡的器官病理变化和脾脏、法氏囊的组织超微结构改变.结果发现人参皂苷及其衍生物组的器官病变程度明显低于病毒对照组,法氏囊和脾脏的超微结构病变轻于病毒对照组,说明人参皂苷及其衍生物在体内可以通过保护机体免疫器官来发挥抗病毒作用.     

人参皂苷及其衍生物体内抗鸡马立克氏病毒的病理组织学观察

为探讨人参皂苷及其衍生物体内抗马立克氏病毒的作用机理,试验用马立克氏病毒人工感染雏鸡模型,口服人参皂苷及其衍生物,动态观察雏鸡的组织病理学变化,并比较治疗组和病毒对照组的病变差异.结果人参皂苷和衍生物治疗组的组织结构病变程度均低于病毒对照组.说明人参皂苷及其衍生物可以降低病毒对组织的损害程度,增强雏鸡对病毒的抵抗力.     

马立克氏病毒感染鸡胚成纤维细胞模型的建立

用雏鸡将马立克氏病毒（MDV）血毒复壮,分离发病鸡淋巴细胞并接种于鸡胚成纤维细胞,观察其病变。获得适应鸡胚成纤维细胞（CEF）的MD强毒,通过电镜观察、琼脂扩散实验进一步鉴定病毒,并建立了MDV感染CEF细胞模型,为研究人参皂苷及其衍生物的体外抗病毒作用及其机制奠定基础。     

人参皂苷及其衍生物抗马立克氏病毒的作用

用马立克氏病毒(Marek′s disease virus,MDV)人工感染雏鸡为疾病模型,采取人参皂苷及其衍生物口服,通过间接免疫荧光试验动态观察人参皂苷及其衍生物能否减少MDV抗原在组织中的分布。结果人参皂苷组、衍生物组被检组织的阳性细胞数量明显比盐酸吗啉胍阳性对照组被检组织的阳性细胞数量少。表明人参皂苷及其衍生物抗MDV效果要优于盐酸吗啉胍。     

人参总皂苷及其衍生物抗Ⅰ型马立克氏病毒作用的电镜观察

应用透射电镜负染技术，观察人参总皂苷及其衍生物对Ⅰ型马立克氏病毒（MDVI）作用后病毒颗粒结构改变情况，旨在深入探讨人参总皂苷及其衍生物抗MDV的作用机理。结果显示，病毒颗粒变形，大小不均匀．核心缺损或均质化，包膜裸露或破损以及病毒颗粒凝集融合成块，结构模糊不清。在相同条件下衍生物比人参总皂苷对MDV的破坏作用强。提示人参总皂苷及其衍生物在体外对MDVⅠ（MDV—Ⅰ）病毒颗粒的形态结构、表面成分和分散均有破坏作用，而衍生物作用更加显著。     

人参皂苷及其衍生物对鸡胚成纤维细胞最大安全浓度的测定

采用组织细胞培养方法,测试了人参皂苷及其衍生物在鸡胚成纤维细胞(CEF)体外培养中的最大安全浓度。结果显示:药物在高浓度时,对CEF细胞表现出毒性,在中、低浓度时对CEF细胞具有促增殖作用。人参皂苷及其衍生物对CEF细胞的最大安全浓度为分别为375μg/mL和187.5μg/mL。     

企业

正大华农取得一项疫苗发明专利证书广东大华农动物保健品股份有限公司8月21日发布公告称,公司近日收到国家知识产权局颁发的一项发明专利证书,发明名称为"一株马立克氏病病毒疫苗株及其分离鉴定和应用"。大华农称,该发明在健康鸡场中分离到一株鸡马立克氏病病毒(MDV),命名为CVTR株。MDV CVTR株不会诱发鸡的肿瘤,对鸡群也没有免疫抑制作用。CVTR株病毒在鸡胚成纤维细胞(CEF)上或鸡体内比目前广泛使用的CVI988/Rispens疫苗株增殖更迅速。将CVTR株用作马立克氏病活疫苗的生产毒株所制备的疫苗,预防超强毒MDV诱发的鸡马立克氏病,其保护力优于目前国内外市场上应用     

MDCC—MSB1培养基上清液中DNA活性的定性研究

将 Ｍ Ｄ Ｃ Ｃ Ｍ Ｓ Ｂ１ ４８ 小时培养物 １０００ｒ／ｍ ｉｎ 离心的上清液分别用 Ｒ Ｎ Ａ 酶、 Ｄ Ｎ Ａ 酶和蛋白酶 Ｋ 处理后，进行体外试验。结果表明，只有 Ｄ Ｎ Ａ 酶处理后的上清液失去了体外抑制 Ｍ Ｄ Ｖ“８１４”增殖的作用。将该上清液 １００００ｒ／ｍ ｉｎ 离心所得的沉淀分别用上述酶处理后进行体内试验。结果表明，只有 Ｄ Ｎ Ａ 酶处理的样品失去了体内促进 Ｍ Ｄ Ｖ 京１ 株致瘤的作用。同时，电泳分析结果证明，该上清液中确实存在 Ｄ Ｎ Ａ。     

An enzyme-linked immunosorbent assay for the detection of antibodies to Marek's disease virus

A reproducible enzyme-linked immunosorbent assay (ELISA) using Marek's disease virus (MDV)-infected cells for the detection of antibodies to MDV is described. The optimum number of MDV-infected chicken embryo fibroblasts (CEF) was 5 X 10(4)/well, and test sera were positive at 1:400 dilutions. Compared with a purified virus preparation, MDV-infected CEF produced high specific and low nonspecific reactivities. Wells coated with whole cells could be stored at 4 C or -20 C for at least 3 months without loss of reactivity. With antibody-negative sera, the cutoff absorbency was 0.20 units. The ELISA was 20-to-40-fold more sensitive than indirect immunofluorescence. Homologous combinations of antisera in wells coated with CEF infected with different MDV serotypes were more reactive at higher dilutions than were heterologous combinations. The procedure described is specific and suitable for large-scale screening of both chicken and monoclonal antibodies against MDV.     

Isolation and Identification of a Field Strain of Marek's Disease Virus and Analysis of Major Pathogenic Genes

In a certain area of Shandong province, Marek's disease (MD) occurred in diseased chickens that had been vaccinated by turkey herpesvirus.In order to isolate the virus strain and detect the virus pathogenicity, agar diffusion test, cell culture and indirect immunofluorescence assay (IFA) were used to isolate the Marek's virus from chicken's blood and feather marrow.The isolated strain was adapted to grow in chick embryo fibroblasts (CEF).Genes involved in pathogenesis of MDV, such as meq, pp38 and 132 bp repeat sequence were amplified by PCR.The obtained sequences were compared with that of standard strains published in GenBank by DNAStar software.The results showed that pp38 gene of the SDAU-1 shared homology from 100% with standard virulent sequence.Analysis of 132 bp repeat sequence and meq gene sequences of the viral genome showed that the isolated virus belongs to the highly virulent MDV strains.     

一株鸡马立克氏病病毒的分离鉴定及主要致病基因分析

山东省某地区鸡马立克氏病疫苗免疫鸡群暴发马立克氏病(MD),为分离得到致病毒株,检测其致病性,采用琼脂扩散试验、细胞培养和间接免疫荧光试验(IFA)等方法从发病鸡的血液及羽髓中分离到一株适应鸡胚成纤维细胞(CEF)生长的马立克氏病病毒。采用PCR方法扩增分离毒株的meq、pp38、132bp重复序列等病毒致病相关基因,所得序列用DNAStar软件与GenBank上登录的参考毒株进行比对分析。结果显示,该分离株SDAU-1的pp38基因与标准强毒序列同源性为100%,132bp重复序列的拷贝数及meq基因的变异均符合MDV强毒株的序列特征。     

Adaptation of Marek's disease virus to the Vero continuous cell line

Marek's disease virus (MDV) is a highly infectious, cell-associated oncogenic herpesvirus. Production of MD vaccines has been limited to primary chicken and duck embryo fibroblast (CEF and DEF) cultures. These have a limited life span and cannot be readily stored in liquid nitrogen. Moreover, the need to prepare CEF and DEF cells on a regular basis from 10 to 11 day-old embryos derived from a flock that must be tested continuously for the presence of avian pathogens adds to the cost of vaccine production. A continuous cell line that would support MDV replication could have significant advantages for the rapid large-scale preparation of MD vaccines. In this report, we describe the adaptation to growth of CEF-grown preparations of serotype 1 and serotype 3 (herpesvirus of turkeys; HVT) strains of MDV in cells of the Vero continuous cell line. Although both viruses produced typical CPE, higher levels of infectious progeny and more extensive virus-specific immunofluorescence were obtained for HVT than for the serotype 1 virus. PCR and pulsed field electrophoresis (PFE) analysis of the DNA from Vero cells infected with either virus confirmed the presence of virus-specific DNA.     

Isolation of serotype 2 Marek's disease virus from birds belonging to genus Gallus in Japan

Serotype 2 of Marek's disease virus (MDV) was isolated from apparently healthy birds belonging to genus Gallus that had no history of vaccination with MDV or herpesvirus of turkeys (HVT). Buffy-coat cells from these birds were inoculated onto chicken embryo fibroblast (CEF) cultures for primary isolation. Thirteen isolates from one golden pheasant and three white silky fowls, three black silky fowls, three Japanese long crowers, and three Japanese bantams produced herpes-like cytopathic effects (CPE) in the CEF cultures. Using serotype-specific monoclonal antibodies to MDV and HVT, 11 isolates were identified as serotype 2 MDV by indirect fluorescent antibody tests. The other two isolates were complicated with serotypes 1 and 3 of MDV-related viruses. Of 13 isolates, three cloned by the limiting-dilution method were further characterized as serotype 2 MDV biologically, genetically, and serologically. The results showed that the birds of the genus Gallus were naturally infected with serotype 2 MDV. This is the first report ever published about the distribution of serotype 2 MDV among healthy birds of the genus Gallus.