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1.
用感染水貂阿留申病病毒G株(ADV-G)的猫肾传代细胞(CRFK)建立了检测水貂阿留申病病毒抗体的PPA-ELISA法。该方法敏感性高于CIEP 16倍,具有快速,准确的优点,可用于病原定位,是水貂阿留申病检疫和研究较为理想的方法. 相似文献
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《中国兽医学报》2019,(11)
为探究水貂阿留申病毒(ADV)VP2主要抗原表位在水貂阿留申病血清学诊断中的作用,本试验将VP2蛋白主要抗原区的表位基因用柔性肽串联连接,将其克隆到表达载体pET-30a(+)中进行原核表达,并对诱导条件进行优化。利用Ni-NTA His-Bind纯化系统对融合蛋白进行纯化,经SDS-PAGE和Western blot鉴定表达产物,并将融合蛋白作为检测抗原进行CIEP试验。结果显示,融合蛋白在IPTG终浓度为1 mmol/L、37℃诱导表达4 h,蛋白表达量最高,其大小约为30 000,以可溶性表达形式为主;融合蛋白能与6×His单克隆抗体及ADV多克隆抗体发生特异性反应,说明该蛋白具有较好的反应原性;且该蛋白作为对流免疫电泳技术(CIEP)检测抗原与ADV阳性血清之间产生了明显的沉淀线,证实了该蛋白作为CIEP检测抗原的可能性。以纯化后的重组蛋白为抗原初步建立的间接ELISA方法具有较高的准确性。结果表明该蛋白具有良好的血清学检测价值,为水貂阿留申病血清学诊断方法的建立奠定了基础。 相似文献
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本实验研究依据阿留申病毒(ADV)的特点,研制了四种不同类型的免疫生物制剂,并分别对临床健康的阿留申病对流免疫电泳(AD.CIEP)阴性水貂接种,攻毒后进行循环免疫复合物(CIC)和抗体(Ab)滴度检测,安死后检查病理组织学变化。本实验结果表明:四种不同类型的生物制剂,其免疫应答反应有显著不同;而同种类型免疫生物制剂,由于剂型、接种剂量不同,其免疫效果亦有明显差异。实验得出,白油佐剂ADV—G亚单位疫苗和白油佐剂(846)抗原的免疫效果好,是用于免疫预防水貂AD有希望的免疫生物制剂。 相似文献
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水貂阿留申病,又称水貂浆细胞增多症。是由病毒引起的一种慢性贫血性传染病。到目前为止尚无特异性治疗方法。国外集中研究特异性诊断方法,以通过检疫、淘汰等措施,逐步控制和清除该病。对流免疫电泳(CIEP)诊断阿留申病得到推广和实际应用。该法特异性强,检出率高,同时适宜大规模检查用。我们于1983年首次在国内研究成功应用 CIEP 检测水貂阿留申病。1986年应用该法俭测,我国饲养的不同基因型水貂,对其发病率进行探讨,以期为我国彩色水貂抗病育种和水貂新色型培育提供依据。现将试验情况报告如下。 相似文献
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水貂阿留申病是水貂的主要传染病之一,每年给水貂养殖业造成约30%左右的经济损失。本病至今尚无特效的防治办法。用阿留申病病毒诊断抗原检出病貂并予以淘汰,是目前世界上控制和扑灭本病的唯一有效办法。农牧渔业部动物检疫所经过一年的反复试验,已于1985年11月成功地研制出阿留申病病毒——猫肾传代细胞诊断抗原。该抗原克服了水貂脏器抗原和猫肾原代细胞抗原存在的非特异性干扰大,动物来源困难、制备 相似文献
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农牧渔业部动物检疫所阿留申病课题组 《中国动物检疫》1986,(1):24-26
水貂阿留申病是水貂的主要传染病之一,每年给水貂养殖业造成巨大的经济损失。本病至今缺乏特异性防治方法。用对流免疫电泳试验逐年检测貂群,严格剔出阳性病貂,保留阴性水貂以作繁殖,是日前世界上防制本病,净化阿留申病阳性水貂场的最有效办法。农牧渔业部动物检疫所采用猫肾传代细胞(CRFK)和阿留中病毒Utah-1国际标准毒株,在国内首次研制成功阿留申病病毒传代细胞抗原。这种抗原具有成分单纯、特异性强、沉淀线清晰、检出率高和成本低、方法简便、适于大批量生产等特点,与国外同类产品相比,结果完全一致,是口岸检疫和貂场净化的理想试剂。 相似文献
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《中国预防兽医学报》2015,(8)
为建立一种适合于基层快速检测水貂阿留申病毒(ADV)的检测方法,本研究将ADV的VP2基因经重叠延伸PCR法进行扩增后克隆至表达载体p GEX-4T-1中进行原核表达,将纯化的重组蛋白作为包被抗原建立了ADV的Dot-ELISA检测方法,并与对流免疫电泳(CIEP)作对照试验。结果表明,抗原最适包被浓度为20μg/m L,最适血清稀释度为1∶200,酶标二抗最适工作浓度为1∶2 000,血清和酶标抗体最适反应温度为37℃,反应时间为45 min~60 min。阻断试验和交叉试验显示与常见的貂病毒性肠炎和貂犬瘟热阳性血清无交叉反应,表明其具有良好的特异性。采用建立的Dot-ELISA和CIEP对78份临床血清样品进行检测,结果显示Dot-ELISA阳性检出率为84.6%,CIEP的阳性检出率为80.8%,敏感性高于CIEP,两者符合率为96.2%。本研究建立的Dot-ELISA检测方法简便、快速、灵敏、特异、重复性好,适合基层单位用于水貂阿留申病快速诊断和疫病普查。 相似文献
8.
水貂阿留申病(Aleutian Disease)在我国已普遍流行,感染率甚高,目前无疫苗及药物治疗,只能通过检测手段来净化貂群。目前在国际上采用的CIEP检验方法,因成本高,条件复杂,不易在国内推广。本文提出了一种快速、检出率高、易掌握、便于推广的酶标斑点纸条法。该法是一种新发展起来的免疫学新技术,其原理是用过碘酸钠化学交联方法将酶分子交联到水貂阿留申病的免疫球蛋白分子上,通过底物的反应作用,生成肉眼可见的颜色反应。此方法优于通常诊断水貂阿留申病的对流免疫电泳法(CIEP)和碘反应法,其特点是:1.灵敏度高,特异性强;2.设备简单,方法简便,标本持久;3.成本低廉,适于批量检验,便于普及推广。 相似文献
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Detection of antibody in Aleutian disease of mink: comparison of enzyme-linked immunosorbent assay and counterimmunoelectrophoresis 总被引：1，自引：0，他引：1
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. 相似文献
12.
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13.
A Hossain Farid 《Acta veterinaria Scandinavica》2013,55(1):10
Background
Aleutian mink disease virus (AMDV) is widespread among ranched and free-ranging American mink in Canada, but there is no information on its prevalence in other wild animal species. This paper describes the prevalence of AMDV of 12 furbearing species in Nova Scotia (NS), Canada.Methods
Samples were collected from carcasses of 462 wild animals of 12 furbearing species, trapped in 10 NS counties between November 2009 and February 2011. Viral DNA was tested by PCR using two primer pairs, and anti-viral antibodies were tested by counterimmunoelectrophoresis (CIEP) on spleen homogenates.Results
Positive PCR or CIEP samples were detected in 56 of 60 (93.3%) American mink, 43 of 61 (70.5%) short-tailed weasels, 2 of 8 (25.0%) striped skunks, 2 of 11 (18.2%) North American river otters, 9 of 85 (10.6%) raccoons, and 2 of 20 (10.0%) bobcats. Samples from six fishers, 24 coyotes, 25 red foxes, 58 beavers, 45 red-squirrels and 59 muskrats were negative. Antibodies to AMDV were detected by CIEP in 16 of 56 (28.6%) mink and one of the 8 skunks (12.5%). Thirteen of the mink were positive for PCR and CIEP, but three mink and one skunk were CIEP positive and PCR negative. Positive CIEP or PCR animals were present in all nine counties from which mink or weasel samples were collected.Conclusions
The presence of AMDV in so many species across the province has important epidemiological ramifications and could pose a serious health problem for the captive mink, as well as for susceptible wildlife. The mechanism of virus transmission between wildlife and captive mink and the effects of AMDV exposure on the viability of the susceptible species deserve further investigation. 相似文献14.
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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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
16.
阿留申病抗体消长规律的研究 总被引：1，自引：0，他引：1
应用阿留申病对流免疫电泳法，历时３年，对正常饲养的貂群逐月进行检测，揭示了ＡＤ抗体在水貂体内的消长规律，提出了适合我国国情的，以ＡＤ．ＣＩＥＰ法一年一次性普检ＡＤ貂的最佳时机，并首次发现了大批ＡＤ抗体阳性貂的长期阴转现象，从而为本病的免疫预防的可能性提供了科学依据。 相似文献
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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。 相似文献
19.
JIA Yun LI Yan-wu SUN Ming-ying ZHANG Xue ZHANG Yan-yan SONG Da-he LIU Zhao WANG Quan-kai SUN Ying-jie 《中国畜牧兽医》2016,43(6):1597-1603
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. 相似文献
20.
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. 相似文献