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水貂阿留申病防制的研究进展 总被引：1，自引：0，他引：1
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。 相似文献
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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。 相似文献
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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。 相似文献
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《中国兽医学报》2014,(12)
为了确定鉴定阿留申病的适宜方法,尤其在阿留申病毒高感染貂场,本试验联合应用对流免疫电泳(CIEP)和PCR方法,对1个貂场连续2年于1112月间检测1次貂血液的阿留申病毒及其抗体。用CIEP检测产仔(断乳成活数≥5只/窝)母貂及其仔貂。通过比较貂群的检测结果来分析2种检测方法鉴别阿留申病貂的适用性。2次检测结果显示各貂群CIEP阳性率为60%12月间检测1次貂血液的阿留申病毒及其抗体。用CIEP检测产仔(断乳成活数≥5只/窝)母貂及其仔貂。通过比较貂群的检测结果来分析2种检测方法鉴别阿留申病貂的适用性。2次检测结果显示各貂群CIEP阳性率为60%97%,PCR阳性率为20%97%,PCR阳性率为20%63%,水貂的CIEP与PCR检测阳性结果不完全匹配。根据CIEP和PCR联合检测结果,貂群被分为4个群体CIEP+PCR+、CIEP-PCR-、CIEP-PCR+和CIEP+PCR-。2次检测的CIEP+PCR-貂在当年母貂中分别占48%、39%,在老母貂中分别占43%、77%,在公貂中分别占21%、51%。2次检测的CIEP+PCR-貂在貂群所占比例大,而CIEP-PCR-和CIEP-PCR+貂在貂群中所占比例小。公貂、老母貂经PCR阳性淘汰间隔1年再次检测,CIEP+PCR-貂在群体中比例明显提高。产仔263%,水貂的CIEP与PCR检测阳性结果不完全匹配。根据CIEP和PCR联合检测结果,貂群被分为4个群体CIEP+PCR+、CIEP-PCR-、CIEP-PCR+和CIEP+PCR-。2次检测的CIEP+PCR-貂在当年母貂中分别占48%、39%,在老母貂中分别占43%、77%,在公貂中分别占21%、51%。2次检测的CIEP+PCR-貂在貂群所占比例大,而CIEP-PCR-和CIEP-PCR+貂在貂群中所占比例小。公貂、老母貂经PCR阳性淘汰间隔1年再次检测,CIEP+PCR-貂在群体中比例明显提高。产仔24年龄母貂有72%4年龄母貂有72%80%CIEP+貂,而其60日龄仔貂仅有11%80%CIEP+貂,而其60日龄仔貂仅有11%16%CIEP+貂。结果分析指出,中国阿留申病病毒高感染率养殖貂群不适宜单纯采用CIEP检测淘汰阳性貂净化阿留申病。发现每年11至12月用CIEP和PCR联合检测貂血样,易检出CIEP+PCR-的潜在阿留申病毒感染康复貂。 相似文献
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水貂阿留申病基因检测芯片的研究与初步应用 总被引：6，自引：0，他引：6
根据已发表的水貂阿留申病病毒（ADV）的序列，设计合成能扩增VP2基因片段的一对引物，通过生物素标记PCR技术，将VP2基因片段作为探针点在硝酸纤维素膜上，制作成疾病诊断基因芯片。以采取的160份可疑病貂的血液核酸作模板，进行PCR扩增，将其产物与诊断基因芯片进行特异性逆向点杂交检测；同时使用水貂阿留申病DOT-ELISA检测试剂盒进行检测、比较。结果表明基因芯片的检出率比ELISA方法高17％，试验同时还比较了常规PCR与基因芯片的敏感性。 相似文献
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为了有效地控制水貂阿留申病的发生 ,保护养貂业的健康发展 ,提高养貂的经济效益 ,用对流电泳免疫法进行了水貂阿留申病的检测。1 材料与方法被检血清 :采自某养貂场自留种貂 440只 ,其中母貂 35 0只 ,公貂 90只。貂趾尖毛细血管采血 ,2 0 0 0～ 30 0 0 rpm离心 2～ 5 min,分离血清备用。抗原 :采用猫肾传代细胞培养的阿留申病毒国家标准毒株 vtah- 1研制而成 ,专供水貂阿留申病对流免疫电泳实验诊断用。由农业部动物检疫所提供。阿留申病阳性和阴性参考血清 ,用于对照 ,由农业部动物检疫所提供。琼脂糖凝板的制备 ,取 3mm厚 1 0× 9cm的普… 相似文献
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Aleutian disease (AD), caused by the Aleutian mink disease virus (AMDV), is a major health concern that results in global economic losses to the mink industry. The unsatisfactory outcome of the culling strategy, immunoprophylaxis, and medical treatment in controlling AD have urged mink farmers to select AD resilient mink based on several detection tests, including enzyme-linked immunosorbent assay (ELISA), counterimmunoelectrophoresis (CIEP), and iodine agglutination test (IAT). However, the genetic analysis of these AD tests and their correlations with pelt quality, reproductive performance, packed-cell volume (PCV), and harvest length (HL) have not been investigated. In this study, data on 5,824 mink were used to estimate the genetic and phenotypic parameters of four AD tests, including two systems of ELISA, CIEP, and IAT, and their genetic and phenotypic correlations with two pelt quality, five female reproductive performance, PCV, and HL traits. Significances (P < 0.05) of fixed effects (sex, year, dam age, and color type), covariates (age at harvest and blood sampling), and random effects (additive genetic, permanent environmental, and maternal effects) were determined under univariate models using ASReml 4.1 software. The genetic and phenotypic parameters for all traits were estimated under bivariate models using ASReml 4.1 software. Estimated heritabilities (±SE) were 0.39 ± 0.06, 0.61 ± 0.07, 0.11 ± 0.07, and 0.26 ± 0.05 for AMDV antigen-based ELISA (ELISA-G), AMDV capsid protein-based ELISA, CIEP, and IAT, respectively. The ELISA-G also showed a moderate repeatability (0.58 ± 0.04) and had significant negative genetic correlations (±SE) with reproductive performance traits (from −0.41 ± 0.16 to −0.49 ± 0.12), PCV (−0.53 ± 0.09), and HL (−0.45 ± 0.16). These results indicated that ELISA-G had the potential to be applied as an indicator trait for genetic selection of AD resilient mink in AD endemic ranches and therefore help mink farmers to reduce the adverse effects caused by AD. 相似文献
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Counter current immunoelectrophoresis (CCIE) is the diagnostic method used in the ongoing Aleutian disease virus eradication program on Danish mink farms. There has been an increasing demand for an alternative diagnostic test especially to evaluate suspected false positive CCIE reactions. We compared test results of a number of negative and positive mink sera in indirect counter current immunoelectrophoresis (ICCIE), counter current line absorption immunoelectrophoresis (CCLAIE) and radio immuno assay (RIA) with test results from counter current immunoelectrophoresis and found that counter current line absorption immunoelectrophoresis is the best alternative diagnostic screening test to counter current immunoelectrophoresis for Aleutian disease eradication programs. Not only proved the CCLAIE test to be useful for evaluation of doubtfully positive CCIE reactions, but it was found to have a higher sensitivity than the CCIE test. 相似文献
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为研究水貂阿留申病毒(AMDV)在体外诱导猫肾细胞(CrFK)凋亡情况,用AMDV-G株感染CrFK细胞,采用CCK-8法检测CrFK细胞感染后的细胞存活率,用AO/EB染色法检测CrFK细胞核的形态变化,用Annexin V-FITC/PI双染流式细胞术检测细胞凋亡率,并通过分光光度计法检测细胞凋亡执行分子Caspase-3活性。结果显示,AMDV-G株感染可导致CrFK细胞增殖率下降,产生明显的形态学凋亡特征;流式细胞术检测结果显示,AMDV-G株感染可引起CrFK细胞凋亡并随着感染时间的延长凋亡率增加,同时Caspase-3活性显著升高。上述结果表明,AMDV-G株在体外能够诱导CrFK细胞凋亡,为进一步研究AMDV致病机理奠定基础。 相似文献
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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. 相似文献
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ZHANG Lei CHAI Xiu-li FENG Pei-ping ZHANG Hai-ling HU Bo BAI Xue ZHAO Jian-jun FENG Zhuo WANG Zhen-jun XU Lei YAN Xi-jun WU Wei 《中国畜牧兽医》2015,42(7):1686-1691
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. 相似文献
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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。 相似文献
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Background
Aleutian disease in mink is caused by infection with Aleutian mink disease virus (AMDV). In Sweden, the infection most commonly causes classical Aleutian disease in which the immune system fails to neutralize the virus and the infection becomes persistent. Diagnosis of AMDV infection is based on serological methods that detect virus-specific antibodies. Traditionally counterimmunoelectrophoresis (CIEP) has been the preferred method, but in order to enable automation interest has been paid to other antibody detecting systems. Recently, at least two different ELISA systems that detect antibodies to AMDV have been manufactured; one is based on an in vitro grown AMDV as antigen, and the other system is based on the AMDV capsid protein VP2 as antigen. The aim of this study was to evaluate the two ELISA systems for detection of antibodies to AMDV using CIEP as the gold standard.Results
When employing the mean optical density of the samples from CIEP negative mink plus three standard deviations as cut-off value, the ELISA with the VP2 antigen had a sensitivity of 99.7% and a specificity of 98.3% compared to CIEP (n = 364). Analysis of samples with the AMDV-G antigen based ELISA employing an assay cut-off value based on the negative control samples, as suggested by the manufacturer, resulted in a sensitivity of 54.3% and a specificity of 93.2% with reference to CIEP as the gold standard (n = 359). When employing the mean optical density of the samples from CIEP negative mink plus three standard deviations as cut-off value, the AMDV-G ELISA had a sensitivity of 37.6% and a specificity of 98.3%.Conclusions
The ELISA system based on VP2 antigen had high sensitivity and specificity, and was concluded to be an alternative to the CIEP as a diagnostic tool for AMDV antibodies. In contrast, the AMDV-G ELISA suffered from low sensitivity when compared to CIEP. 相似文献20.
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。 相似文献