水貂阿留申病病毒CIEP抗原结合蛋白初探

以水貂阿留申病病毒对流免疫电泳(CIEP)细胞抗原为材料,经酶印迹(Westemblotting)测定,水貂阿留申病病毒CIEI细胞抗原与多克隆阳性血清反应,分子量为60000,50000和25000,而与CIEP阴性的抗水貂阿留申病病毒的单克隆抗体(Y—2—9)反应,分子量为60000,50000.因此初步确定水貂阿留申病病毒CIEP细胞抗原决定族位于分子25000蛋白上.     

水貂阿留申病基因工程诊断抗原的制备

本研究旨在建立一种生产水貂阿留申病(Aleutian disease,AD)诊断抗原的新方法。试验提取水貂阿留申病毒(Aleutian disease virus,ADV)的基因组,PCR扩增ADV核衣壳蛋白VP2基因,构建重组表达质粒Bacmid-VP2,脂质体介导其转染昆虫细胞Sf9获得重组杆状病毒AcMVPV-VP2。电镜下观察表达的VP2蛋白,Western blotting检测目的蛋白的反应原性。以传统接毒方法生产的AD诊断抗原作对照,通过对流免疫电泳试验检测表达蛋白的生物学活性。结果表明,表达的重组VP2蛋白在电镜下组装成病毒样颗粒且能与ADV阳性血清发生反应。与商业诊断抗原相比,重组抗原诊断AD的阴阳性的符合率为100%。该方法可成为生产AD诊断抗原的替代方法。     

Preparation of Genetic Engineering Diagnostic Antigen of Mink Aleutian Disease

To develop a new method of Aleutian disease (AD) diagnostic antigen production,we used Bac-to-Bac expression system in this study.Firstly,Aleutian disease virus (ADV) genome was extracted and ADV VP2 gene was amplified by PCR method.Then Bacmid-VP2 was constructed and transfected into insect cell Sf9 by liposomes to construct AcMVPV-VP2.Secondly,the VP2 protein was observed by electromicroscope and antigency was detected by Western blotting.At last,the activity of recombinant protein was inspected by countercurrent immunoelectrophoresis.The results showed that the expressed recombinant VP2 protein could react with ADV positive serum and form virus like particles.Compared with the commercial diagnostic antigen,the coincidence rate of recombinant antigen was 100%.This method could be a candidate for AD diagnostic antigen production.     

Ultraviolet Inactivation of the Infective Agent of Aleutian Disease of Mink

An experiment carried out to examine the effect of ultraviolet light on the aleutian disease agent in serum indicated that the agent was sensitive to irradiation.     

水貂阿留申病的研究进展

水貂阿留申病(Aleutian disease of mink,ADM)是由水貂阿留申病细小病毒(Aleutian mink disease parvovirus,AD-MV)引起的一种慢性、进行性传染病,一直是危害世界养貂业健康发展最重要的疫病之一。到目前为止,还没有疫苗可成功用于ADM的预防,也没有特异有效的治疗方法,唯一可行的防治方法就是通过多次特异性检疫,淘汰病貂,净化貂群。笔者对阿留申病的病原学、发病机制、防治措施等方面进行概述,为临床防治水貂阿留申病提供了理论基础。     

Clinical Chemical Studies in Aleutian Disease of Mink

Clinical chemical determinations were carried out on blood removed by cardiac puncture from 49 mink affected with Aleutian disease and 25 normal animals and the respective differences tested for statistical significance. Blood urea nitrogen, serum total protein and globulin, thymol turbidity, glutamic oxalacetic and glutamic pyruvic transaminases and amylase were definitely elevated in the affected animals whereas serum calcium, albumin and A/G ratio were depressed. No statistically significant difference was apparent between the two groups in the comparison of inorganic phosphorus, alkaline and acid phosphatases, bilirubin, total cholesterol and esters, cephalin-cholesterol flocculation (3+ in each case), sodium, potassium, chloride, CO₂-combining power, leucine aminopeptidase and lactic dehydrogenase (means: over 2,000 u./ml.). For both the control and affected mink, the distribution of serum lactic dehydrogenase isozymes resembled that of human homologous serum hepatitis. Electrophoresis of serum proteins confirmed earlier findings of hypergammaglobulinemia in the diseased animals but a fast-moving or pre-albumin component, averaging 4% of the total protein, occurred in both the diseased and normal mink.     

水貂阿留申病灭活疫苗免疫效果观察

为评价水貂阿留申病灭活疫苗的免疫效果 ,对接种疫苗水貂及阿留申病阴性、阳性水貂的死亡、空怀、流产、产仔、产仔成活数进行了比较 ,结果证实 ,水貂阿留申病灭活疫苗对水貂具有较好的免疫保护作用。     

Aleutian Disease (Plasmacytosis) of Mink III. Propagation of the Virus in Mink Tissue Cultures

Cultures of mink testis and mink kidney cells were inoculated with 10% extracts and filtrates of tissue from plasmacytosis-affected mink. Specific morphological changes were observed in cultures of kidney and testis cells. Millipore filtration experiments suggested the size range of the agent to be from 10 to 50 mμ. Inoculation of fluids from the 2nd, 4th and 8th tissue culture passages of the agent induced plasmacytosis in adult mink.     

水貂阿留申病毒的分子生物学研究进展

从水貂阿留申病毒(ADV)基因组特点出发,就阿留申病毒的分子生物学研究进展作以简单综述。     

水貂阿留申病毒实验室检测方法研究进展

水貂阿留申病是一种在世界范围内广泛流行的、严重危害水貂养殖业的病毒性免疫抑制性传染病。本文介绍了水貂阿留申病毒的分离与鉴定、血清学和分子生物学最新实验室检测方法研究进展,以期为防控本病提供参考。     

对流免疫电泳在水貂阿留申病诊断中的应用进展

水貂阿留申病(AD)是一种在世界范围内广泛流行的、严重危害水貂养殖业的病毒性传染病。对流免疫电泳(CIEP)是世界公认的诊断AD病的有效方法。本文综述了CIEP在AD诊断中的应用进展,介绍了该方法的优缺点和实用性,以期为疾病的有效防控提供参考。     

Aleutian Disease (Plasmacytosis) of Mink: II. Responses of Mink to Formalin-Treated Diseased Tissues and to Subsequent Challenge With Virulent Inoculum

An experiment was carried out to examine the responses of Aleutian and standard dark types of mink to inoculations of formalintreated suspensions of tissues of mink with experimentally transmitted plasmacytosis. Control groups of mink received similar injections of normal Aleutian mink tissues or diseased tissues without formalin treatment. A second experiment was conducted to test the formalinized diseased tissue suspension for immunogenic value. Groups of mink which received one, two, or three doses of “vaccine” were later challenged with virulent inocula. Additional groups of mink served as unvaccinated and environmental controls.

Treatment with 0.3% formalin with fine trituration and incubation at 37°C was effective in preventing the development of plasmacytosis in inoculated mink. These mink remained susceptible to subsequent challenge with untreated diseased tissue suspensions. No immunity was demonstrated in the vaccinated mink. Mink inoculated with normal mink tissues did not develop plasmacytosis, nor did uninoculated environmental controls.

     

水貂阿留申病毒结构蛋白与非结构蛋白的研究进展

水貂阿留申病毒(Aleutian mink disease virus,ADV)是一种主要侵染水貂的自主复制型细小病毒,是一种在水貂中广泛存在的重要病原体。病毒粒子的蛋白分为结构蛋白(VP1、VP2)和非结构蛋白(NS1、NS2)两类。VP1蛋白对病毒粒子产生感染性有重要作用;VP2蛋白是主要免疫功能区,能刺激机体产生中和抗体;NS1和NS2主要参与病毒的复制和基因的表达调节。文中对近年来国内外学者关于水貂阿留申病毒结构蛋白和非结构蛋白的研究情况进行归纳和总结。     

Myocardial Necrosis and Mineralization in Normal and Aleutian Disease Mink Fed Dexamethasone

Focal myocardial necrosis with mineralization occurred in mink fed the synthetic corticosteroid, dexamethasone, as a treatment for aleutian disease. Cardiac lesions occurred with equal frequency in steroid-fed controls and appeared to be related only to feeding of dexamethasone. Possible pathogenetic mechanisms are considered.     

Aleutian Disease of Mink: I. Evidence of its Viral Etiology *

A suspension of tissues from field cases of Aleutian disease was used successfully to reproduce the disease in Aleutian mink. Similarly, suspensions of diseased tissues from the experimentally infected mink were used to transmit the agent of Aleutian disease to both Aleutian mink and standard dark mink. Seitz and millipore filtrates prepared from these tissue suspensions were also infective; a suggestion that the etiologic agent is a virus. Genetic factors and hypersensitivity are discussed as possibly contributing to development of the disease.     

水貂阿留申病毒部分VP2基因的原核表达及免疫学分析

利用PCR技术扩增出水貂阿留申病毒(ADV)含有VP2抗原表位区的基因片段,将其克隆到原核表达载体pGEX-4T-1中,构建出重组质粒pGEX-4T-VP2,并转化入大肠杆菌BL21中,用IPTG法进行诱导表达。经SDS-PAGE凝胶电泳和免疫印迹分析显示有目的条带,并且在诱导6 h后表达量达到最高,随时间的延长表达量降低。本研究成功构建了pGEX-4T-VP2重组质粒,确定了VP2基因的优化表达条件,为阿留申病的免疫学诊断和疫苗研制奠定基础。     

Studies on Viral Plasmacytosis (Aleutian Disease) of Mink : VII. Infection of Mink with DNA Extracted from Diseased Spleens

Lesions considered typical of aleutian disease developed in three of four mink inoculated with DNA extracted from spleens of mink with viral plasmacytosis. Control mink inoculated with buffered saline or DNA treated with specific enzyme (DNAase) remained normal. It is inferred that the infective DNA corresponds to viral DNA.

This DNA preparation was used in an attempt to infect tissue cultures from mink testis cells but the results were equivocal.

     

水貂阿留申病的历史回顾及最新研究概述

水貂阿留申病是阻碍养貂业发展的世界性攻关病害。自人们发现该病至今已有５０余年的历史,尽管国内外有关专家学者,从没间断过对该病的研究,但始终没能攻克该病的免疫防制问题;值得庆幸的是,在国家科委和农业部畜牧兽医司等部门的大力资助下,我们对阿留申病的免疫原性和免疫机制进行了较系统深入的研究,并研制出了阿留申病灭活疫苗,现已在多个大型貂场进行小试。该文对阿留申病的病史和危害,以及最新研究进展和重大技术突破进行了较全面系统的论述。     

水貂阿留申病毒结构蛋白与非结构蛋白在疾病诊断与疫苗研发中的应用

水貂阿留申病是由水貂阿留申病毒引起的一种持续感染性疾病,危害毛皮动物养殖业的发展.水貂阿留申病毒的致病特点、免疫机制等与其他细小病毒不同,存在自身的复杂性.目前,众多研究者寻求新的疫苗来预防水貂阿留申病的发生,但没有取得理想的效果.疾病诊断及疫苗研发对防控水貂阿留申病具有积极的意义.水貂阿留申病毒结构蛋白在病毒感染、机...