水貂阿留申病毒检测方法研究进展

水貂阿留申病是由水貂阿留申病毒引起的以浆细胞增多为主要特征的慢性、消耗性、病毒性传染病。主要侵害水貂的免疫系统,导致机体免疫系统紊乱引起的器官衰竭,病死率极高,严重损害了水貂养殖产业的经济利益,该病存在于绝大多数养殖水貂的国家和地区。目前,能有效预防该病的疫苗尚未研制成型,该病的防控已成为水貂养殖领域的世界性难题,在防控方面主要采取淘汰检疫阳性水貂的办法。论文主要针对目前该病在国内外检测方法的特点和流行情况进行阐述,以期为水貂阿留申病快速检测方法的建立及水貂阿留申病的有效防控提供参考。     

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...     

某貂场暴发疑似水貂阿留申病的诊断报告

水貂阿留申病(AD)是由阿留申病毒(ADV)引起水貂的一种慢性、进行性传染病。以病毒持续性感染,典型的获得性免疫缺陷综合症为特征。国外对本病研究报告较多,国内尚缺乏系统研究。查见山西、吉林对水貂饲养场进行了调查,但未见貂场有如此凶猛的流行,仅从一些侧面反映了该病的流行情况,而我省未见报道。1989年7月至11月在我地某水貂饲养场发生了一次疑似水貂阿留申病的流行,现将诊断结果报告如下:     

水貂阿留申病的普查

自1946年美国学者确定阿留申病为一种独立性疾病以来,在美国、苏联、挪威、瑞典、芬兰、丹麦、德国到波兰等养貂业比较发达的国家中都有本病流行的报道,而且发病的水貂大批死亡不能出口。近年来,国内有些水貂场也有该病的发生,且有蔓延的趋势。虽然阿留申病被公认为一种病毒性疾病,但也可能存在阿留申病的遗传基团,故阿留     

板蓝根液治疗水貂阿留申病

水貂阿留申病对养貂业危害极大。该病流行面广、发病率和死亡率高,严重影响水貂繁殖机能。笔者用以下方法共治疗12例病貂,全部治愈。1流行特点该病病原为阿留申病毒,呈垂直传播和水平传播,秋冬季发病和死亡率高,经感染的水貂大部分病例为慢性经过,病期为数周至数月。2临床症状患貂表现为食欲减退,进行性消瘦,口渴暴饮,可视黏膜苍白,齿龈出血和溃疡,排煤焦油样粪便。3剖检变化该病主要侵害肝、脾、肾等实质器官,死后常见这些器官肿大和灰白坏死灶。4治疗方法根据流行特点、临床症状、剖检变化等可诊断为水貂阿留申病。对病貂应用板蓝根注射液…     

我国首次分离出水貂阿留申病病毒(ADTVC_4)强毒株、并应用金黄色葡萄球菌菌体协同凝集反应诊断水貂阿留申病

水貂阿留申病病毒的分离、鉴定及特异性诊断方法的研究成果于1983年10月10日在哈尔滨通过鉴定。本课题的研究是由东北林学院野生动物系和黑龙江省兽药一厂共同主持进行的。阿留申病毒是水貂阿留申病的病原体。该病是一种慢性进行性传染病,在欧洲、美洲、亚洲等二十多个养貂国家普遍流行,此病在我国呈蔓延趋势。     

水貂阿留申病的诊断与防治

<正>本文综述了水貂阿留申病的不同诊断方法,介绍了水貂阿留申病目前常用的防治措施,这有助于该病的诊断及提前预防,提高经济效益。水貂阿留申病又称浆细胞增多症或高丙球蛋白血症。这种病是由细小病毒科,细小病毒属的阿留申病病毒(ADV)侵染机体引起的,经发展成为一种慢性的持续性衰弱疾病。患病机体常表现为终生病毒血症,持续性感染,单核—巨噬细胞系统受到侵害。其生理特征是浆细胞弥漫性增生、进行性免疫复合物形成、产生多量γ球蛋白,     

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。     

板蓝根治疗水貂阿留申病

<正> 水貂阿留申病对养貂业危害极大,该病流行面广,发病率和死亡率高,严重影响水貂的繁殖机能。笔者用以下方法共治疗12例病例,全部治愈,现将结果报告如下。1 流行特点该病的病原为阿留申病毒,呈垂直传     

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.     

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。     

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。     

阿留申病细小病毒的分离及VP2基因遗传衍生分析

水貂阿留申病（Aleutian disease of mink，AD）是水貂的一种慢性传染病，病原为阿留申病细小病毒（Aleutian mink disease parvovirus，AMDV），属细小病毒科、细小病毒属。AD自1940年发现以来至今60年里，已经普遍存在于世界各地人工养殖的水貂种群中。对水貂养殖业造成了不可估量的经济损失。     

Nucleotide sequence and polymerase chain reaction/restriction fragment length polymorphism analyses of Aleutian disease virus in ferrets in Japan.

Two ferrets with spontaneous Aleutian disease (AD) were found in Japan. The diagnosis was verified by polymerase chain reaction (PCR) amplification of part of the capsid gene specific to AD virus (ADV). The nucleotide sequences (365 bp in length) of the amplified fragments from the 2 ferrets differed by a single nucleotide, producing an amino acid alteration. Compared with other types of ADV, these isolates had 96% sequence similarity to a published ferret ADV (FADV) in contrast to <91% homology to various types of mink ADV (MADV). The phylogenetic tree of ADVs indicates that these 2 isolates and the published FADV belong to the same genetic group and definitely are divergent from MADVs. The predicted amino acid sequence of the hypervariable segment in the capsid gene was conserved among the 3 types of FADV. These results indicated that the 2 isolates found in Japan were new DNA types of FADV and could have been derived from FADV(s). A restriction fragment length polymorphism (RFLP) method to distinguish the ferret types of ADV from the mink types of ADV was developed on the basis of differences in their nucleotide sequences. Digestion of the PCR products with Afal or ScaI provided different cleavage patterns for FADV and MADV. This PCR/RFLP analysis of the ADV capsid gene will be a valuable asset for diagnosis of this virus infection in ferrets.     

Characteristics of inapparent Aleutian disease virus infection in mink.

Inapparent of nonprogressive Aleutian disease virus (ADV) infection is a subclinical but persistent virus infection of mink. Mink with the inapparent type of ADV infection when subjected to stress did not develop the progessive form of the disease. However, when challenged with a large dose of the virus, these mink did develop progressive Aleutian disease indicating that they were not highly resistant to the virus. Sera of mink with either the progressive of the inapparent type of ADV infection did not neutralise the virus. The anti-ADV antibody activity in mink with inapparent type of ADV infection was in the IgG fraction of the serum the same as in mink with progressive Aleutian disease. These data indicate that the resistance of the mink with inapparent infection as compared to mink with progressive Aleutian disease was not due to a difference in the class of immunoglobulin response to the virus. However, mink with progressive Aleutian disease showed a greatly increased immunoglobulin response.     

Detection of antibody in Aleutian disease of mink: comparison of enzyme-linked immunosorbent assay and counterimmunoelectrophoresis

Experiments were undertaken to investigate the potential of the enzyme-linked immunosorbent assay (ELISA) as a screening test for the diagnosis of the 2 known naturally occurring forms of Aleutian disease of mink. Anti-Aleutian disease virus (ADV) antibody activity was not detectable in the sera of mink with nonprogressive Aleutian disease despite the demonstration of antibody by counterimmunoelectrophoresis (CIEP) in the same sera. Anti-ADV antibody was detectable in 93% of sera from mink at various stages of experimentally induced progressive Aleutian disease. False-negative reactions occurred in sera which demonstrated high anti-ADV antibody titers by CIEP. As a consequence of the high prevalence of false-negative reactions, the ELISA was not considered to be an effective screening test. However, using CIEP as an indicator of ADV infection, the ELISA may be useful in differentiating mink with nonprogressive Aleutian disease from mink with progressive Aleutian disease.     

Mink infected with aleutian disease virus have an elevated level of CD8-positive T-lymphocytes

Lymphocytes, monocytes, granulocytes, B-lymphocytes and CD8-positive T-lymphocytes of non-infected mink and mink infected with Aleutian disease virus (ADV) were measured by flow cytometry. The gammaglobulin levels of the sera were also measured. Besides development of hypergammaglobulinaemia in the infected mink, the most pronounced finding was that the number of CD8-positive lymphocytes doubled on average during development of Aleutian disease, while the number of B-lymphocytes did not change dramatically. The enhanced CD8 frequency was still apparent 6 months after initial ADV infection of the mink. The present experiments contribute to a better understanding of the immune deficiency stage seen in mink infected with ADV.     

Mechanisms contributing to the virus persistence in Aleutian disease

In this review published results and further studies concerning the persistence of Aleutian disease virus (ADV) isolate SL3 are presented. By Southern blot and in situ hybridization with strand-specific RNA probes focal replication of ADV-DNA was demonstrated in spleen, mesenteric lymph nodes, sporadically in mononuclear cells of the peripheral blood and bone marrow cells. These findings further support the concept of the lymphotropism of ADV. All cell culture-adapted ADV strains appear to have a ts-defect. Our in vitro studies indicate that the ADV isolate G(orham) induced the synthesis of comparable amounts of viral replicative DNA and viral proteins VP1 and VP2 at the non-permissive temperature of 37 degrees C. However, the viral progeny DNA synthesis was about threefold less at 37 degrees C compared to the permissive temperature of 32 degrees C. These findings suggest that the reduced level of viral progeny DNA at 37 degrees C accounts for the reduced production of infectious ADV. Finally, we provided experimental evidence that the apparent lack of neutralizing antibodies in AD is due to the masking of critical viral epitopes by cellular phospholipids.