水貂阿留申病诊断技术研究进展

水貂阿留申病是水貂养殖业的重要疫病,目前无疫苗预防,主要通过发展诊断技术,淘汰感染貂防控该病。论文总结了在貂场应用的基因与抗体检测技术,基因检测具有高灵敏度和特异性,能够检测到动物带毒感染,不同类型的基因检测技术对变异性强的阿留申病毒检测各有所长;抗体检测是水貂阿留申病的主要检测方式,研究者不断创新和改进原有技术,趋向对阿留申病的规模化精准检测,但单一的抗体检测方法淘汰感染貂具有局限性。论文通过对两类检测技术分析,建议采用基因与抗体的联合检测方法,从感染貂群中区分耐受貂或抗性貂,期望为国内高感染率貂场的检测和选种提供参考。     

水貂阿留申病的诊断与体会

<正>水貂阿留申病是由阿留申病毒引起的一种慢性、接触性传染病。本病的主要特点是潜伏期长、浆细胞增多、血丙种球蛋白含量增高、抗原抗体复合物肾小球肾炎、进行性衰竭等。潜伏期水貂大多不表现临床症状,遇有饲养条件恶劣、气候寒冷潮湿等情况可导致病情恶化,引起水貂死亡。水貂阿留申病可引起发育不良,皮张质量降低,繁殖力下降,母貂空怀,流产,仔兽成活率降低,是阻碍养貂业发展的重大疫病之一。1发病情况     

水貂养殖场对流免疫电泳检测阿留申病注意事项

正水貂阿留申病是目前危害水貂养殖业的三大疫病之一,该病可明显降低水貂产仔率、仔貂成活率、皮毛质量以及疫苗免疫后的抗体水平。当前,国内一些大型水貂养殖场或者是种貂场为了净化水貂阿留申病,提高生产业绩,每年进行2次(打皮期、分窝期)或者3次(打皮期、配种期、分窝期)水貂阿留申病     

水貂阿留申病对流免疫电泳用诊断抗原的制备和试用

本文报道从金州水貂场的银蓝貂中分离出一株水貂阿留申病毒(ADV),定名为《辽金81-02》株。用它人工感染8月龄健康水貂并连续传代,可以保持病毒的强毒力。收获急性感染9天后发病貂的脾、肝及淋巴结作为种毒材料加以结冻保存,并成功地用它们制取阿留申病毒对流免疫电泳抗原,用于检测抗阿留申病毒的抗体,诊断疾病。 制取病毒抗原的方法:首先是用氟炭提取含毒组织乳剂,再经重复超速离心加以浓缩及部分提纯,并用盐酸-甘氨酸处理进行活化。每批抗原要用对照抗原及阳性血清进行抗原活性鉴定。 用这种初步纯化的活性抗原,能在水貂感染后7天,在对流免疫电泳中检出阿留申病毒的沉淀性抗体。试验证实:用对流免疫电泳检测阿留申病毒和抗体是特异性的,这要比碘凝集试验的特异性和敏感性高得多。经过对比试验,我们所制10批抗原的特异性、复制性及敏感性均可和美国的商品抗原相比。电泳时,可在45分钟内,在1％琼脂糖凝胶板的抗原抗体孔之间出现清晰可见的沉淀线。 Cho及Greevfield已报告,可以成功地用对流免疫电泳试验从病貂群中清除阿留申病。我们用试制抗原在金州水貂场对1500只各色型水貂进行AD抗体检测,结果扑杀阳性貂所提示的典型阿留申病的病理病变是符合一致的;与美国同类商品抗原平行检测130只貂的结果,也是符合     

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。     

水貂阿留申病的防控措施

正水貂阿留申病广泛流行于世界各养貂国家,我国各养貂场均有此病发生,本区的水貂养殖场水貂阿留申病都有不同程度的感染,2014年,本区两水貂养殖场部分水貂,经对流免疫电泳检测,阿留申阳性率分别达到87.8%(36/41)和77.3%(17/22)。水貂阿留申病的危害很大,严重影响毛皮动物的繁殖能力,免疫机能和毛皮质量。1水貂阿留申病的流行特点     

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...     

水貂常发病的防治

水貂主要有阿留申病、病毒性肠炎和犬瘟热三大疫病，为有效控制水貂这三大疫病的发生，保护养貂业的健康发展，提高水貂的产仔率和成活率，提高经济效益，笔者在去年进行了水貂疫病综合防治技术推广，并列入开发区“丰收计划”项目，先后进行了对流电泳免疫法检测水貂阿留申病、细胞苗防疫水貂病毒性肠炎和大瘟热试验，这一实验项目，今年元月经专家鉴定验收合格，并达国内先进水平。l对流电泳免疫法检测水貂阿留申病试验1．l试验时间与工也点1998年1月10日至10月10日在黄岛街道办事处柳沟村养貂场进行试验。1．2材料与方法1．2．1被检血清…     

水貂阿留申病的危害及防控对策

水貂阿留申病是由阿留申病毒感染水貂引起的以终生毒血症、全身淋巴细胞增殖、肾小球肾炎为特征的一种慢性病毒病。各品系水貂都可以感染,是世界公认的水貂的三大疫病之一。该病既可以通过飞沫、粪便、尿液、血液、配种等水平传播,又可以通过母婴垂直传播。     

水貂阿留申病对犬瘟热及细小病毒性肠炎疫苗免疫的影响

对81份水貂血样进行了水貂阿留申、犬瘟热及细小病毒性肠炎的抗体进行检测,并对检测结果进行了对比分析,表明阿留申抗体阳性水貂的犬瘟热及细小病毒性肠炎抗体合格率、均值,均远低于阿留申抗体阴性貂,验证了阿留申病对水貂的免疫应答有较为明显的影响。     

Transmission of Aleutian disease from mink with inapparent infections.

In apparent or nonprogressive Aleutian disease virus infection was considered a subclinical but persistent viral infection in which infected mink did not develop tissue lesions, hypergammaglobulinemia, or high antibody titers. Transmission of Aleutian disease virus from mink with this type of infection was measured. Mink with inapparent Aleutian disease appeared healthy and had normal gamma-globulin values, but were capable of transmitting the disease by direct and indirect horizontal contact. The risk of direct or indirect horizontal transmission from mink with inapparent infection was less than from mink with progressive Aleutian disease. Infection also was directly transmitted from the dam to the kits, but again the risk of infection from dams with inapparent infection was less than from dams with progressive Aleutian disease. Mink infected from their dams before weaning developed the disease more slowly than mink which became infected after weaning.     

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.     

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.     

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.     

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。     

Isolation and Identification of Local Aleutian Mink Virus Virulent Strain ADV-LN

Mink suspected infection aleutian mink disease virus (ADV) from mink breeding areas in Liaoning province were tested with CIEP method.The mink with antibody to ADV were selected and culled.Liver,spleen,kidney and mesenteric lymph node samples were taken for pathological examination and the viruses were observed under electron microscope.The grinded tissue fluid filter was added penicillin and treptomycin and inoculated into CRFK cells and passaged by 6 times for virus isolation.And cells cultures were identified as ADV by PCR.Then they were inoculated into healthy mink.Three days later,the mink showed clinical signs,which including the loss of appetite,anemia,hair dull,antifeedant and binge drinking.Some minks showed neurological symptoms,manifested symptoms of convulsions,cramps,staggering gait,ataxia,or hind limb paralysis and died.The virus strains isolated and identified were named as the ADV-LN.     

水貂阿留申病毒地方强毒株ADV-LN的分离鉴定

本研究采用对流免疫电泳方法(CIEP)检测辽宁水貂养殖场疑似水貂阿留申病毒(aleutian mink disease virus,ADV)水貂血清抗体,采集抗体阳性水貂的肝脏、脾脏、肾脏和肠系膜淋巴结组织,电镜观察存在细小病毒样颗粒。组织液研磨无菌处理后,接种CRFK细胞,盲传6代,取病毒细胞分离液用PCR方法检测,呈ADV阳性。将病毒分离液纯化后接种健康水貂,隔离观察,接种后3 d即出现食欲减退,贫血,被毛无光泽,后期出现拒食、狂饮、死亡,个别水貂出现神经症状,表现抽搐、痉挛、步态蹒跚、共济失调,证明分离获得的病毒为ADV强毒株,命名为ADV-LN株。     

Mink with Aleutian disease have autoantibodies to some autoantigens

Thirty mink infected with Aleutian disease virus (ADV) were found to have elevated levels of antibody to double-stranded DNA (dsDNA) in their sera when compared to 30 healthy mink. The anti-dsDNA antibody levels in the diseased mink were, however, not found to correlate with the total amount of immunoglobulin. This was a common observation for all autoantibodies tested. The concentration of rheumatoid factors of IgG class, but not those of IgM class, was found to be significantly higher in the diseased mink at the chosen level of significance (P less than 0.01). IgG antibodies to thyroglobulin were likewise significantly higher in the ADV-infected mink. Unexpectedly, we found IgG antibodies with specificities for cardiolipin and mitochondrial antigens to be significantly higher in healthy mink than in ADV-infected mink. This difference is especially remarkable since the serum immunoglobulin concentration of the ADV-infected mink was three times higher than the serum immunoglobulin concentration of the normal mink.     

Detection of inapparent Aleutian disease virus infection in mink.

The normal serum gamma-globulin centration of mink from the Ontario Veterinary College field station was 13.2 +/- 2.6% of total serum proteins. Mink serum gamma-globulin concentrations above 21%, which represented 3 standard deviations above the normal mean, were considered to be hypergammaglobulinemic. About 39% of pastel mink infected naturally with Aleutin disease virus (ADV) exhibited an inapparent or nonprogressive infection. These nonprogressivley infected mink had serum gamma-globulin values below 21% andhad antibody titers less than 256 if tested by the couterimmunoelectrophoresis technique. Mink maintained inapparent infection for at least 10 months after infection with ADV. Neither gross nor histopathologic changes were present in the mink with inapparent ADV infection. The virus persisted in blood, mesenteric lymph nodes, kidney, liver, and spleen of mink with non-progressive infection, although the amount of virus present probably was small.     

Isolation of virus and antibody containing immune complexes from mink with Aleutian disease by affinity chromatography of equine complement clq

Affinity chromatography on immobilized equine complement Clq was used for the isolation of complement-binding immune complexes in sera of mink infected with Aleutian disease virus. Immune complexes were isolated and quantitated from 4 of 5 infected mink, as early as 2 weeks after infection and before hypergammaglobulinemia had appeared. The quantity of immunoglobulin G in these immune complexes ranged from 180 to 370 micrograms/ml serum. There were no Clq-binding immune complexes found in mink which were negative for Aleutian disease antibody. Using 125I-labeled BSA-anti-BSA complexes, we demonstrated that the affinity columns bound selectively immune complexes which had formed in antibody excess, whereas immune complexes in antigen excess were not bound. By neutralization of sensitized virus with anti-mink IgG serum, non Clq-binding immune complexes were also detected, which indicates that circulating immune complexes in persistently infected mink are heterogeneous as far as their reactivity with equine Clq is concerned.