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1.
免疫胶体金快速诊断技术源于对血清与尿液的绒毛膜促性腺激素的免疫渗滤试验检测,之后有学者提出采用胶体金作为标记物替代酶应用于渗滤试验,并制备完成广泛应用的对抗人类免疫缺陷病毒抗体检测的试剂盒.由于免疫胶体金快速诊断技术方便快捷,已在妊娠试验、检测传染病病原抗体等方面进行应用,而应用于兽医诊断的研究刚刚起步. 相似文献
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ELISA技术及其在禽病诊断中的应用 总被引：1，自引：0，他引：1
ELISA,在1971年最初建立时称为酶联免疫吸附剂测定(enzyme linked immunosorbent assay),在国内有人译作酶联免疫吸附试验,它是一种免疫测定,是把抗原抗体反应的特异性和酶的高效催化作用有机结合起来的一种非放射性标记免疫检测技术。近年来,随着单克隆抗体技术和酶标记技术的发展与应用,市场上已出现了各种各样的商品化ELISA试剂盒,广泛应用于临床医学、动植物食品检验、生物学研究等领域。本文主要是对ELISA技术的基本原理、类型、影响因素及其目前在禽病诊断中的应用加以综述。1ELISA技术简介1.1ELISA基本原理ELISA是以免疫… 相似文献
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电化学酶联免疫传感器在农畜产品安全检测中的应用研究 总被引：2，自引：0，他引：2
电化学酶联免疫传感器是一种标记型的免疫传感器,它结合了酶电极的放大作用和免疫传感器的灵敏性及特异性,是近几年来研究最多、发展最快、应用最广的一种免疫传感器。综述了电化学酶联免疫传感器的特点及其在农畜产品安全领域（农药残留、兽药残留等）的研究进展和应用现状,并对电化学酶联免疫传感器的发展前景进行了展望。 相似文献
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现代诊断技术在兽医学上的应用及其进展 总被引：2，自引：0，他引：2
1 免疫酶技术 (ＥＩＡ)ＥＩＡ是根据抗原 -抗体结合的特异性 ,以酶作标记物 ,酶对底物具高效催化作用的原理而设计 ,既可用于抗原的检查 ,也常用于血清抗体的检测。1 .1 酶联免疫吸附试验 (ＥＬＩＳＡ)ＥＬＩＳＡ是应用最广泛的免疫酶技术 ,包括间接法、夹心法、竞争法等。自ＶＯＬＬＥＲ和ＢＩＤＷＥＬＬ(1 975 )首先报道了应用间接ＥＬＩＳＡ方法检测病毒特异性抗体以来 ,ＥＬＩＳＡ方法已普遍应用于传染病的流行病学普查、抗体水平的评价以及疫苗质量的监控和标化等 ,迄今为止大多数动物病原体均已发展或报道了ＥＬＩＳＡ方法的研究与… 相似文献
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周永华 《上海畜牧兽医通讯》1992,(2):38-40
近年来,免疫酶技术以其高度的特异性和敏感性,在弓形虫病诊断和流行病学调查中得到了广泛应用,从而推动了弓形虫病的防治研究进入了一个新阶段。本文就几种常用的免疫酶方法在弓形虫病的应用概述如下: 一、酶联免疫吸附试验(ELISA) 1971年Engvall和Van Weeman等首次创立ELISA技术。其基本原理就是用酶催化底物反应的生物放大作用,提高抗原抗体免疫学反应的检测敏感性。Volley等首先将ELISA用于检测弓形虫病患 相似文献
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免疫酶技术的研究进展很快,目前在国外逐渐形成高潮,着重研究对某些畜禽病毒、细菌、支原体及寄生虫感染的诊断,从当前的研究结果来看,免疫酶技术具有和荧光抗体法同样的敏感性、特异性及快 相似文献
8.
免疫胶体金技术是一种新型免疫标记技术,与免疫荧光技术、酶联免疫吸附技术和放射免疫测定法相比更具优越性。本文综述了胶体金技术的基本原理、制备方法、标记技术以及目前在禽病诊断中的应用情况。 相似文献
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ELIB诊断黄牛东毕血吸虫病的研究 总被引：1，自引：1，他引：0
本研究应用酶联免疫印清技术分析了东毕血吸虫成虫部分纯化抗原中具有免疫活性的蛋白质组分，其中分子量为３５、８０、９２和９４ｋＤ的蛋白质具有较灵敏的反应原性，可做为东毕血吸虫病的诊断抗原。本试验应用此种抗原探索了诊断黄牛东毕血吸虫病的方法，取得了满意结果。 相似文献
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Development and evaluation of an indirect enzyme immunoassay for detection of porcine antibodies to pseudorabies virus. 总被引：3，自引：1，他引：2 
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下载免费PDF全文 An indirect enzyme immunoassay is described for detection of porcine serum antibody to pseudorabies virus. The analytical sensitivity of the enzyme immunoassay was found to be approximately 4.5 log 4 X 10 (5120 times) greater than the serum neutralization test, based on parallel end point titrations. The diagnostic sensitivity of the enzyme immunoassay was comparable or superior to that of the serum neutralization, based on the earliest detectable antibody after infection of swine with pseudorabies virus by intranasal or intrauterine routes or by contact with infected pigs. The enzyme immunoassay, at a screening dilution of 1:20, gave 100% agreement with ELISA results provided with a U.S. Department of Agriculture-Animal and Plant Health Inspection Service proficiency panel of 40 sera. One serum having demonstrable antibody by the enzyme immunoassay was seronegative by the serum neutralization test. 相似文献
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通过比较孔雀 Ig G与鸡 Ig G的相似性 ,证实鸡疾病诊断试剂用于孔雀疾病诊断的可行性。用饱和硫酸铵盐析法提纯鸡 Ig G、孔雀 Ig G,比较 Ig G粗提物的 SDS-聚丙烯酰胺凝胶电泳图谱。用直接 EL ISA方法分析孔雀 Ig G、鸡 Ig G与抗鸡 Ig G结合程度的差异 ,并用蛋白免疫转印法进一步验证二者与抗鸡 Ig G的结合情况。最后 ,应用数种鸡疾病诊断试剂盒及血凝抑制实验进行孔雀疾病的血清学调查及诊断应用初步研究。结果表明 :孔雀 Ig G与鸡 Ig G具有较大相似性 ,并能够与兔抗鸡酶标二抗结合 ,但结合力弱于鸡 Ig G;可应用鸡疾病诊断试剂盒诊断孔雀疾病 相似文献
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W A Fleenor D O Lucas G H Stott A J Guidry 《Veterinary immunology and immunopathology》1984,6(3-4):365-378
Monoclonal antibodies were developed to bovine IgG1. In addition, production of monoclonal antibodies to bovine light chain is also reported. Monoclonal antibody specificities were initially determined by solid-phase enzyme immunoassay. The monoclonal antibovine IgG1 was shown by a specificity-independent isotyping solid-phase enzyme immunoassay to be mouse IgG1 with kappa light chains. Ascites derived monoclonal antibovine IgG1 antibodies were linked to cyanogen bromide-activated Sepharose and used for affinity isolation of bovine IgG1. The bovine IgG1 eluted from the affinity column was characterized using immuno-electrophoresis, acrylamide gel electrophoresis, isoelectric focusing and solid-phase enzyme immunoassay. Affinity chromatography using monoclonal antibodies provided both a verification of monoclonal antibody specificity and a rapid technique for the isolation of bovine IgG1. This technique may also be employed to remove IgG1 contaminants during purification of bovine IgA. 相似文献
15.
Enzyme-linked immunosorbent assay for differentiation of the antibody response of cattle naturally infected with Brucella abortus or vaccinated with strain 19 总被引：10，自引：0，他引：10
K Nielsen J W Cherwonogrodzky J R Duncan D R Bundle 《American journal of veterinary research》1989,50(1):5-9
Purified O chain of Brucella abortus was passively attached to polystyrene to differentiate antibody responses of cattle vaccinated with B abortus strain 19 from those of naturally infected cattle. In the indirect assay, using O polysaccharide as antigen, a single serum dilution was used and mouse monoclonal antibody to bovine L chain conjugated with horseradish peroxidase was the detection reagent. Measurable antibody was not found in sera of vaccinated cattle, except for 3 sera from cattle that were persistently infected with strain 19. Sera from 25 cattle infected with pathogenic strains contained antibody on the basis of results of indirect enzyme immunoassay, using smooth lipopolysaccharide or O chain as antigens, or results of competitive enzyme immunoassay, using the O-chain antigen. Results in sera from calves with experimentally induced Yersinia enterocolitica serotype 0:9 infection or inoculated with a low dose of B abortus strain 2308 were comparable with those in sera of cattle that were vaccinated with strain 19. The data correlated with those from competitive enzyme immunoassay, using one serum dilution and horseradish peroxidase-conjugated mouse monoclonal antibody to smooth lipopolysaccharide. On the basis of results of the indirect enzyme immunoassay, all sera (except those samples obtained before inoculation) contained antibody to smooth lipopolysaccharide. 相似文献
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S T Grubbs S A Kania L N Potgieter 《Journal of veterinary diagnostic investigation》2001,13(2):123-127
Subgroup-specific peptide-based enzyme immunoassays from each respective G-glycoprotein of the ovine and the bovine respiratory syncytial virus (RSV) were developed to detect RSV-specific IgG responses in cattle. Antigenic peptides from the respective G-glycoprotein were identified from the extracellular central hydrophobic region (amino acids 158-189) located between 2 mucin-rich regions. These antigenic peptides identified by epitope mapping from each G-glycoprotein were synthesized and used to develop the subgroup-specific enzyme immunoassays. The negative cutoff for each enzyme immunoassay was established as the mean optical density of indirect immunofluorescent antibody-negative bovine sera plus 3 SDs. The sensitivity (82.9%) and specificity (100%) of the bovine enzyme immunoassay and the specificity (95.8%) of the ovine enzyme immunoassay were determined by comparison with indirect immunofluorescence (used as the "gold standard"). The negative and positive predictive values were calculated for each assay. The presence of serum antibody to ovine RSV in cattle implies that this virus infects cattle and may contribute to the pathogenesis of bovine respiratory disease. 相似文献
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Validation of the fluorescence polarization assay as a serological test for the presumptive diagnosis of porcine brucellosis. 总被引：5，自引：0，他引：5
K Nielsen D Gall P Smith A Vigliocco B Perez L Samartino P Nicoletti A Dajer P Elzer F Enright 《Veterinary microbiology》1999,68(3-4):245-253
Sera from Canadian pigs (brucellosis free, n = 14037) and sera from pigs infected with Brucella suis (n = 401) were tested by the buffered antigen plate agglutination test, the complement fixation test, an indirect and a competitive enzyme immunoassay and a fluorescence polarization assay. The results were analysed and assay sensitivity and specificity estimates were calculated. The sensitivity and specificity of the tests were as follows: the buffered antigen plate agglutination test, 77.1 and 96.9%; the complement fixation test (considering anticomplementary sera as negative), 93.3 and 95.5%; the complement fixation test (considering anticomplementary sera as positive), 58.1 and 99.9%; the indirect enzyme immunoassay, 94.0 and 97.9%; the competitive enzyme immunoassay, 90.8 and 96.6%; and the fluorescence polarization assay, 93.5 and 97.2%; respectively. It was concluded that the fluorescence polarization assay was a valuable asset to the diagnosis of porcine brucellosis because of its accuracy, ease of performance and relative cost. 相似文献
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Evaluation of an enzyme immunoassay for the detection of antibodies against Sarcocystis spp. in naturally infected cattle in China 总被引：1，自引：0，他引：1
An enzyme immunoassay was used to detect anti-Sarcocystis antibodies in the sera of 159 cattle sent to slaughter in Jilin Province, north China. The musculature of each carcase was closely examined for the presence of parasitic cysts and aliquots of muscle were digested with pepsin and examined microscopically for cystozoites. Specific antibodies were detected in 126 (79.25%) cattle whereas cystozoites were detected in 123 (77.36%) and cysts in 103 (64.78%). The accuracy, sensitivity and specificity of the enzyme immunoassay were high (0.97, 0.99 and 0.91, respectively) and false reactions were only detected in four cases. The assay exhibited a high level of reproducibility when samples were re-tested and the soluble Sarcocystis spp. antigens did not cross-react with anti-Toxoplasma antibodies raised in rabbits. This report presents the first successful application of an enzyme immunoassay in the diagnosis of Sarcocystis spp. infections in naturally infected cattle in China. The assay, however, failed to detect specific antibodies in four dogs experimentally infected with S. cruzi from cattle. 相似文献
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I R Dohoo P F Wright G M Ruckerbauer B S Samagh F J Robertson L B Forbes 《Canadian journal of veterinary research》1986,50(4):485-493
Five serological assays: the buffered plate antigen test, the standard tube agglutination test, the complement fixation test, the hemolysis-in-gel test and the indirect enzyme immunoassay were diagnostically evaluated. Test data consisted of results from 1208 cattle in brucellosis-free herds, 1578 cattle in reactor herds of unknown infection status and 174 cattle from which Brucella abortus had been cultured. The complement fixation test had the highest specificity in both nonvaccinated and vaccinated cattle. The indirect enzyme immunoassay, if interpreted at a high threshold, also exhibited a high specificity in both groups of cattle. The hemolysis-in-gel test had a very high specificity when used in nonvaccinated cattle but quite a low specificity among vaccinates. With the exception of the complement fixation test, all tests had high sensitivities if interpreted at the minimum threshold. However, the sensitivities of the standard tube agglutination test and indirect enzyme immunoassay, when interpreted at high thresholds were comparable to that of the complement fixation test. A kappa statistic was used to measure the agreement between the various tests. In general the kappa statistics were quite low, suggesting that the various tests may detect different antibody isotypes. There was however, good agreement between the buffered plate antigen test and standard tube agglutination test (the two agglutination tests evaluated) and between the complement fixation test and the indirect enzyme immunoassay when interpreted at a high threshold. With the exception of the buffered plate antigen test, all tests were evaluated as confirmatory tests by estimating their specificity and sensitivity on screening-test positive samples.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献