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.     

水貂阿留申病碘凝集与对流免疫电泳检测方法比较

水貂阿留申病是危害水貂养殖业的三大疫病之一,目前没有疫苗可以使用,也没有特效的治疗药物,只能通过检测、淘汰来净化貂群。本研究随机采取河北某貂场准备配种后期的水貂血清样品168个,采用碘凝集和对流免疫电泳进行检测,结果表明:二者的符合率为81.55%。鉴于二者的符合率及检测成本,建议生产中首先用碘凝集对留种貂群进行初筛,再进一步用对流免疫电泳检测,采用二者相结合的方法对水貂种群进行净化。     

水貂阿留申病净化研究

阿留申病是主要侵染水貂的一种病程缓慢的传染性疾病,是危害养貂业的重要疾病之一。为了净化水貂场的阿留申病,从根本上杜绝该病的发生、传播。针对不同饲养条件的水貂场,采用对流免疫电泳检测结合渐进隔离饲养法对种貂群进行净化。结果表明:采用此两种方法,5年内基本完成种貂群阿留申病的净化工作,对母貂的生产性能不但未产生不良影响,其生产性能反而有了显著提高,受胎率、产仔数等均明显提高。     

水貂阿留申病病毒研究进展

水貂阿留申病(Aleutian mink disease, AMD)是由水貂阿留申病病毒(Aleutian mink disease virus, AMDV)引起的水貂的重要传染性疾病之一,深入开展对AMDV的研究对于该病的防控有重要意义。AMDV基因组全长约为4.8 kb,主要编码2种结构蛋白和3种非结构蛋白,它们在病毒复制、增殖及致病过程中发挥重要作用。AMDV的复制依赖于代谢活跃的细胞,对于幼貂,病毒感染肺泡Ⅱ型细胞会造成急性致死性肺炎,感染巨噬细胞则会引起成年水貂患高丙种球蛋白血症和免疫复合物介导的肾小球肾炎等慢性进行性疾病。笔者从AMDV侵入细胞的受体途径、诱导细胞凋亡途径及病毒复制等方面对其致病机理进行阐述。AMDV在全世界范围内广泛流行,现有的检测方法主要分为血清学诊断方法和分子生物学诊断方法。目前,尚未开发出安全有效的针对AMDV的商品化疫苗,随着生物学技术的快速发展,在灭活疫苗、DNA疫苗和亚单位疫苗的研制上有所进展;抗病毒的新方法,如筛选AMDV耐受貂,提高水貂免疫力和靶向适配体技术为AMD的防控提供了新思路。文章从AMDV编码蛋白功能、病毒细胞嗜性与复制、临床表现...     

水貂阿留申病诊断抗原研究概述

近年来,随着水貂养殖行业的不断发展,一些疫病也成为了制约水貂养殖业发展的重要因素。水貂阿留申病作为毛皮动物的三大疫病之一(阿留申病、犬瘟热、病毒性肠炎),是导致母貂产仔率下降、公貂配种能力降低和毛皮质量下降的一种高度接触性传染病。至今为止,还没有商品化的疫苗来控制该病的传播及蔓延。控制水貂阿留申病最好的方法是通过检测淘汰所有抗体为阳性的水貂,进而达到净化貂群的目的。而在抗体检测过程中,诊断抗原的制备和纯化决定着检测方法的敏感性、特异性和准确性。论文对目前阿留申病毒细胞抗原及基因工程抗原研究进展做一综述,为今后该病病原检测工作提供参考。     

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。     

水貂阿留申病防制的研究进展

水貂阿留申病防制是水貂养殖领域的世界性难题。该病临床特征是全身性淋巴细胞增殖、血清γ－球蛋白增高、肾肿大、动脉性血管炎和肝炎、持续性毒血症。多年来,人们对水貂阿留申病防制的研究主要集中于对该病的诊断和淘汰,经历了碘凝集反应,对流免疫电泳技术两个主要阶段。近年来,通过研究阿留申病循环免疫复合物的测定、抗体消长规律、免疫机制、试用免疫接种,使最终攻破阿留申病防制难关出现曙光。     

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。     

Evaluation of chemical disinfectants for aleutian disease virus of mink

Nine chemicals and commercial disinfectants were tested for inactivation of Aleutian disease virus of mink. In the presence of distilled water, a commercial disinfectant (O-Syl), halogen derivatives (iodophor and sodium hypochlorite), and glutaraldehyde (2.0%) inactivated 4 log10 (based on 0.25 ml) of the virus within 10 minutes at 23 C. Formalin (2.0%) and O-Syl were slower to inactivate the virus, but achieved a 4 log10 reduction in titer by 30 minutes' contact time. In the presence of 10% bovine serum, formalin (1.0%), O-Syl, and sodium hydroxide (0.5%) achieved a 4 log10 reduction within 10 minutes. All agents tested had some virucidal effect.     

Poly IC therapy in aleutian disease of mink.

Twenty-four virgin female aleutian mink were infected with aleutian disease agent and after 24 hours, 12 of these were treated with a course of polyinosinic acid-polycytidilic acid (Poly IC) injections. After six weeks the gammaglobulin level was significantly lower in the treated group but at 12 weeks this difference was no longer present. Four of the treated mink had normal target organ histology when killed at 20 weeks. The untreated group all showed moderate to marked changes but this difference was not statistically significant. There was a marked increase in the reactive lymphocyte blastogenesis index during the first weeks of infection and the phytohaemagglutinin response was seen to fall progressively. The antiglobulin reaction usually became positive after infection but neither antinuclear nor antierythrocyte antibodies were found. Precipitating antibodies to several polynucleotides were frequently present and were unrelated to infection or to Poly IC treatment.     

闽北地区猪伪狂犬病抗体检测分析

为了解猪伪狂犬病在闽北地区的流行情况,整理并分析2015年度血清抗体检测数据。采用ELISA检测方法对闽北地区45个规模化猪场进行检测分析,通过检测样品血清中伪狂犬病gE抗体和gB抗体的水平进行结果判定。结果显示：送检的45个规模化猪场,伪狂犬病gE抗体阳性场达25个,猪场阳性率达55.6%;共检测1304份血清,其中伪狂犬病gE抗体阳性213份、可疑24份、阴性1067份,抗体阳性率达20.6%。从伪狂犬病gE抗体阴性猪场抽检猪伪狂犬gB抗体380份,gB抗体阳性258份,猪群伪狂犬病抗体保护率仅有67.9%。     

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.     

Borna disease virus infection in domestic cats: evaluation by RNA and antibody detection

Borna disease virus (BDV) infection has been suggested to cause spontaneous neurological disease in cats referred to as staggering disease. However the evaluation of BDV infection in neurologically asymptomatic cats remained unclear. In the present study, BDV infected, asymptomatic cats in Tokyo were surveyed both by the presence of plasma antibodies against BDV-p24 and -p40 and by RNA detection in peripheral blood mononuclear cells. Seven of 32 domestic cats (21.9%) were serologically or genetically judged to be BDV-infected. Six cats were positive for anti-BDV antibody and two cats were positive for BDV RNA. Within the 2 RNA-positive cats, only one was positive for anti-BDV antibodies. Furthermore, the findings of anti-BDV-p40 and anti-BDV-p24 antibody-positive cats did not completely overlap. These results suggest that there are neurologically asymptomatic domestic cats infected with BDV present in the Tokyo area.     

Single dilution ELISA for detection of serum antibody to foot-and-mouth disease virus in cattle

A single dilution blocking ELISA was developed and evaluated for measuring serum antibody to foot-and-mouth disease virus (FMDV). Basic parameters of the assay were established and a positive-negative threshold determined from testing 176 specific antibody negative sera from Australian cattle. Sera collected from immunised animals in Thailand were tested by ELISA and virus-neutralisation (VN) tests and the results compared. A positive correlation between ELISA and VN titres was recorded for each of the 3 FMDV serotypes endemic in Thailand, with the overall correlation coefficient being r = 0.8990. A positive correlation for each of the serotypes was also found between ELISA titre and the degree of blocking (percentage inhibition) of each test serum at a dilution of 1:16, with the overall correlation being r = 0.8704. This simplified ELISA was sensitive, specific and gave reproducible results, and had the potential to test quickly and efficiently a considerable number of sera.     

兔病毒性出血症病毒抗体间接ELISA检测方法的建立

建立一种检测兔病毒性出血症病毒(RHDV)抗体的间接ELISA方法。对RHDV陕西分离株VP60基因进行原核表达,Western blot分析表达产物的免疫反应性;以纯化的蛋白为包被抗原建立ELISA方法,并对反应条件进行优化。结果表明,VP60蛋白在大肠杆菌中成功表达,产物约为42.34 ku的融合蛋白,具有良好的反应原性;优化的ELISA最佳工作条件为:重组抗原包被浓度2.9μg/mL,37℃2 h后4℃过夜,1%BSA 37℃封闭2 h,待检血清37℃孵育1 h,酶标抗体1∶10 000稀释,37℃作用1 h,37℃显色5 min,临界值为0.340;建立的ELISA方法特异性强、重复性好、敏感性高;临床检测180份样品,与血凝抑制试验的符合率为74.1%,与商品化试剂盒检测结果符合率为94.8%。该方法可用于临床样品的大批量检测。     

猪口蹄疫抗体不同检测方法的比较分析

世界动物卫生组织(OIE)将口蹄疫列为A类疾病,我国将其列为一类传染病,并定为强制免疫病种[1-2].疫苗免疫是预防该病的有效措施之一,如何很好地监测评价疫苗免疫效果是防制该病的关键,同时也为科学制定免疫程序、做好疫病风险评估及考核免疫政策措施落实情况之分析提供依据.