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应用DT试验、IHA Kit与本实验室建立的IHA方法平行检测49份猪血清样品,DT检测出10份阳性样品,10份阳性样品中,IHAKit检出8份阳性,本室建立的IHA方法检出7份阳性.其中DT检测出的39份阴性样品中IHA Kit全部检测为阴性,本室建立的IHA方法检出1份阳性、38份为阴性.同DT试验比较,IHA试验的敏感性、特异性、符合率和Youden指数分别为76.92%、97.5%、91.85%和0.744. 相似文献
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目的:分析ELISA方法和IHA方法检测弓形虫病的可靠性。方法:本文应用酶联免疫吸附试验(ELISA)和间接血凝试验(IHA)两种方法,平行检测来自龙岩市某个猪场的32份猪血清中的抗弓形虫特异性抗体。结果:ELISA和IHA对猪弓形虫抗体阳性检出率分别为62.5%(20/32)、53.13%(17/32),这两种方法的阳性检出符合率较高,为71.88%(23/32)。其中,ELISA方法的敏感性(93.33%)、检测效率(81.25%)以及Youden指数(63.92%)都在IHA方法之上,但ELISA方法的特异性(12/17,70.59%)略低于I-HA方法(13/17,76.47%)。结论:ELISA和IHA两种方法均可用于猪弓形虫病的诊断和血清学调查,但ELISA方法更适用于猪弓形虫病的血清学调查。 相似文献
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用斑点酶联免疫吸附试验(Dot-ELISA)对200份经粪检和间接血凝试验(IHA)血检的血清样品进行肝片形吸虫感染的检测,同时在屠宰场现场对50份样品进行检测,并与剖检相比较。结果显示Dot-ELISA与IHA法相比,阳性符合率较低(84.21%),阴性符合率高(95.58%),总符合率94.50%;Dot-ELISA与粪检结果相比,阳性符合率(94.74%)明显高于IHA法;与剖检法比较,Dot-ELISA对屠宰场现场样品的检出率和阳性符合率均为100%。结论:Dot-ELISA较IHA法敏感性高、特异性强,适宜用作牛、羊肝片形吸虫感染的现场检测。 相似文献
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传染性支气管炎病毒(M41株)HI试验抗原的特异性和敏感性试验 总被引：1，自引：0，他引：1
采用本研究室制备的3批传染性支气管炎病毒(IBV,M41株)HI试验抗原,分别对100份和80份不同鸡血清进行IBV HI抗体检测,同时用IBV ELISA抗体检测试剂盒进行检测,比较两种不同方法检测的特异性和敏感性。结果显示,特异性试验中80份SPF鸡血清,自制抗原检测均为阴性,IBV ELISA检测79份为阴性,10份其他鸡病血清,两种方法检测9份均为阴性,10份IBV阳性血清两种方法检测均为阳性;敏感性试验中,74份已知IB疫苗免疫或IBV M41株感染鸡血清IBV HI检测72份为阳性,阳性检出率为97.3%(72/74),IBV ELISA检测74份均为阳性,阳性检出率为100%(74/74),SPF鸡血清及其他鸡病血清,两种方法检测均为阴性,两种检测方法总符合率为97.5%,差异不显著(P0.05)。试验结果证明,本研究室自制抗原具有良好的特异性,特异性为100%,抗原同时具有良好的敏感性,但IBV ELISA方法的敏感性要高于IBV HI方法。 相似文献
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采用淋巴细胞杂交瘤技术,研制出的抗衣原体单克隆抗体(McAb),应用酶联免疫吸附试验(ELISA)方法,对本地区所采的1361份牛血清,698份羊血清进行检测,结果:牛血清阳性检出率为9.18%,羊为22.06%。选择牛羊血清各50份用已建立的检测衣原体抗体ELISA方法与传统的间接血凝试验(IHA)进行比较,其敏感性高。牛血清ELISA阳性检出率为32%,IHA为24%,ELISA比IHA试验高出8个百分点。羊血清ELISA阳性检出率为26%,IHA为22%,前者比IHA高出4个百分点。 相似文献
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检测2种猫科动物猫瘟热病毒抗体SPA-ELISA方法的建立及初步应用 总被引：1，自引：0，他引：1
利用猫瘟热病毒重组VP2蛋白作为包被抗原,以辣根过氧化物酶(HRP)标记的SPA作为广谱第二抗体,初步建立适用于2种猫科动物猫瘟热病毒抗体检测的SPA-ELISA方法。用建立的SPA-ELISA方法对临床30份猫血清样本、86份虎血清样本进行检测,同时与HI、间接ELISA方法进行比较。结果显示,确定纯化重组VP2蛋白以2.67 mg/L稀释质量浓度包被,脱脂乳以15%的浓度封闭90 min,待检血清以1∶50稀释度孵育60 min,HRP-SPA以1∶4 000稀释度作用45 min,可使SPA-ELISA获得最佳检测效果。SPA-ELISA批内及批间重复试验变异系数均小于10%,对临床30份猫血清样本、86份虎血清样本的对比检测表明,SPA-ELISA方法检出率较高,大于HI法的检出率,与间接ELISA法接近。3种检测方法检测猫血清的总体符合率为96.7%。在虎血清检测中,SPA-ELISA方法的阳性检出率亦高于HI方法,两者总体符合率为94.2%。 相似文献
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通过比较研究羊流产嗜衣原体与弓形虫抗体的间接血凝试验(IHA)与酶联免疫吸附试验(ELISA)检测结果,选择适宜贵州省山羊流产血清学调查的检测方法。通过IHA和ELISA两种方法对贵州省195份山羊血清进行羊流产嗜衣原体与弓形虫抗体的检测,统计并分析两种疫病的阳性符合率、阴性符合率和总符合率。结果表明,羊流产嗜衣原体与弓形虫IHA与ELISA的总符合率分别为77.95%和78.97%,阳性符合率均仅为50%,阴性符合率为80.45%和79.27%。说明在检测两种疫病血清抗体方面,IHA比ELISA更适合在贵州省基层推广。 相似文献
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为了建立猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)抗体检测的间接ELISA方法,本研究以纯化的原核表达的PEDV截短N蛋白作为包被抗原,建立了PEDV抗体检测的间接ELISA方法,将该方法命名为rnPED-ELISA。该抗原不与其他常见的7种猪病的阳性血清发生交叉反应,批内和批间重复性试验的变异系数均小于13%;rnPED-ELISA相对于血清中和试验(SN)试验的敏感性为93.33%,特异性为90.00%;rnPED-ELISA与TSZ全病毒抗体检测试剂盒的符合率达91.67%。采用rnPED-ELISA方法检测200份临床样品,PEDV抗体阳性检出率为69.5%。本试验建立的rnPED-ELISA方法具有良好的敏感性和特异性,可为免疫猪群抗体监测和猪流行性腹泻流行病学调查提供一种快速、简便的血清学诊断方法。 相似文献
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Surya Sankar Hiron M. Harshan S.R. Somarajan S.K. Srivastava 《Research in veterinary science》2010,88(3):375-378
A recombinant leptospiral lipoprotein, LigB, was evaluated for use in the diagnosis of bovine leptospirosis by enzyme-linked immunosorbent assay (rLigB IgG ELISA). The standard reference test (Microscopic agglutination test, MAT) of 200 serum samples from cattle suspected of leptospirosis showed that 95 (47.5%) samples had positive agglutination titres, which ranged from 100 to 1600. In rLigB IgG ELISA, 49% of the samples were positive. Sensitivity of IgG ELISA for 95 bovine sera, which had MAT titres of greater than or equal to 100, were 100%. ELISA showed a specificity of 97.1% with 105 bovine sera, which were negative at a 1:50 dilution in MAT for Leptospira interrogans serovars. The results of ELISA and MAT correspond very good. When analytical specificity of IgG ELISA was evaluated using bovine serum samples from animals showing the serum antibodies to other pathogens, no cross-reaction was observed. Thus the recombinant LigB IgG ELISA can be used instead of the MAT as an aid to the diagnosis of bovine leptospirosis. 相似文献
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Yamanaka T Tsujimura K Kondo T Matsumura T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2008,70(2):189-192
In this study, we evaluated whether five rapid antigen detection kits for human influenza could be used for the diagnosis of equine influenza (EI). Limiting dilution analyses showed that Directigen Flu A+B and ESPLINE INFLUENZA A&B-N had the highest sensitivities to equine-2 influenza viruses (EIVs) among the kits investigated. From the results of virus detection in nasal swabs taken from horses infected with EIV, these two kits could produce positive results in reasonable agreement with those obtained by virus isolation or RT-PCR, suggesting that these kits could be useful for rapid diagnosis of EI in the field. However, from the viewpoint of specificity for EIV, Espline seems to be superior to Directigen. 相似文献
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Comparison of a commercial ELISA with the modified agglutination test for detection of Toxoplasma infection in the domestic pig 总被引：1，自引：0，他引：1
The modified agglutination test (MAT) and a commercially available enzyme-linked immunosorbent assay (ELISA) were compared for detection of antibodies to Toxoplasma gondii in naturally-infected market-aged pigs. Infected pigs were obtained from commercial slaughter facilities and from farms where infection had previously been detected. Infection was confirmed by bioassay in cats. For 70 bioassay positive pigs, 60 were positive by MAT (85.7% sensitivity) and 62 were positive by ELISA (88.6% sensitivity). Of 204 bioassay negative samples 193 were negative by MAT (94.6% specificity) and 200 were negative by ELISA (98.0% specificity). Good correlation was seen between MAT and ELISA results. The results suggest that the ELISA may be a good tool for epidemiological studies of Toxoplasma infection on pig farms. 相似文献
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本试验采用121 ℃高压处理副猪嗜血杆菌4型和5型耐热蛋白,混合作为包被抗原,建立了检测副猪嗜血杆菌抗体的间接ELISA方法。通过对试验条件进行筛选优化,确定了最佳反应条件:抗原包被浓度为10 μg/mL,37 ℃包被2 h;封闭液选择含20 g/L脱脂奶粉的PBST,封闭30 min;血清的稀释度为1∶80;抗原抗体反应时间为45 min;酶标二抗稀释度为1∶12000,作用时间为30 min;底物显色时间为15 min。特异性、重复性和敏感性试验及对200份送检血清的检测结果表明,建立的间接ELISA方法特异性和重复性良好,敏感性比间接血凝试验高,对已知阴阳性血清的临床样本检测结果与国外ELISA试剂盒一致,可用于副猪嗜血杆菌的血清抗体检测和血清流行病学调查。 相似文献
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Kittelberger R O'Keefe JS Meynell R Sewell M Rosati S Lambert M Dufour P Pépin M 《New Zealand veterinary journal》2006,54(1):10-15
AIM: To determine the diagnostic capability of a newly developed Western blot (WB) assay for the detection of serum antibodies against Mycoplasma agalactiae compared with conventional serological tests, and to identify the best test for routine diagnostic use. METHODS: The serological test methods used were: two commercial indirect enzyme-linked immunosorbent assays (ELISA), viz ELISA-1, using a bacterial antigen preparation, and ELISA-2, using a recombinant protein (lipoprotein p48) antigen; the complement fixation test (CFT); and a newly developed WB assay, the latter both using a bacterial antigen preparation. Thirty sera from goats infected with M. agalactiae and 97 sera from non-infected sheep were tested using all four methods. RESULTS: Staining patterns in the WB were quite variable. An immuno-dominant band of 41 kDa was detected in 63% of sera from infected animals. The same band also appeared, although mostly very weakly, in 10% of sera from non-infected animals. When suspicious or very weak reactors were omitted, the diagnostic sensitivity (DSE) and diagnostic specificity (DSP), respectively, for the four assays were: WB=56.7%, 97.9%; ELISA-1=76.7%, 99.0%; ELISA-2=56.7%, 100%; and CFT=40.0%, 94.8%. CONCLUSIONS: ELISA-1 performed best in this comparison. While the WB can be used, it did not have a technical advantage over the ELISA. The CFT should be discouraged as the primary screening method for contagious agalactia and should be replaced by ELISA-1. Results from this study confirm that serological test methods for contagious agalactia are useful for the detection of infected flocks but will not detect every individual infected animal. 相似文献
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An enzyme-linked immunosorbent assay (ELISA) for the detection of IgG antibodies in rodents was developed and validated with the microscopic agglutination test (MAT) and leptospiral cultures. Sonicated antigen from cultures of serovars tarassovi and pyrogenes was used. As conjugate, a combination of anti-rat and anti-hamster IgG labeled with peroxidase was used. The optimal cut-off point was determined by plotting the sensitivity and specificity for various cut-off point values by means of receiver operating characteristic (ROC) curve. Concordance between ELISA and each of the MAT titers was measured by kappa (kappa). Proportions of positive results were compared by means of McNemar's test. Total 214 rodents were trapped, but only 117 could be processed by the three techniques (culture, ELISA, MAT; 1:20, 1:40, 1:50) and used for statistical analysis. Although, MAT titers in rodents infected with the serogroup Ballum tended to be lower than those infected with the serogroup Icterohaemorrhagiae, all (20/20) were ELISA-positive and almost all (19/20) were MAT-positive.The percentage of positive results obtained by ELISA, 47.0% exceeded significantly the 40.2% obtained by MAT (1:50). Difference between ELISA and MAT (1:40) was not significant and no differences were observed between ELISA and MAT (1:20). Agreement, specificity, sensitivity and the consequent area under the ROC curve between ELISA and MAT were higher as MAT cut-off points were lowered, being optimal at 1:20. The fact that differences between ELISA and MAT were significant at 1:50, but not at 1:40 or 1:20, supports the suggestion that lower MAT titers should be considered positive in rodents. The ELISA developed to detect leptospire-specific antibodies had optimal sensitivity and specificity in relation to MAT and it is concluded that it may constitute a very useful indicator for epidemiological purposes of past or present leptospiral infection in rodents. 相似文献
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Leptospirosis is a zoonotic disease with worldwide endemicity in Argentina, it is a significant public health problem in low-income populations. Bovine leptospirosis is a serious economic problem for cattle production, causing abortions, reduced milk yield, mortality in calves and decreased daily weight gain. We developed an enzyme-linked immunosorbent assay (ELISA) with sonicated Leptospira interrogans serogroup Icterohaemorrhagiae serovar Copenhageni M 20. We evaluated its performance for the detection of specific antibodies against multiple Leptospira serogroups in bovine. The microscopic agglutination test (MAT) was used as the gold standard. The performance of this ELISA was evaluated with a panel of sera (118 MAT confirmed positive and 97 MAT negative). The overall sensitivity was close to 85.6 % and the specificity was 83.5 %, according to the MAT reference method. Analytical specificity of the IgG-ELISA was evaluated using 50 bovine serum samples from animals showing serum antibodies against other pathogens that cause abortion in bovine, such as Brucella sp., Neospora sp. and Bovine Viral Diarrhea (BVD), and no cross-reaction was observed. This IgG-ELISA can be an alternative to the MAT for diagnosis of leptospiral infection in bovine. 相似文献
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The efficiency of indirect haemagglutination (IHA) and immunoelectrophoresis (IEP) as means of detection of cystic hydatid infection in camels was investigated. Both IHA and IEP showed very low detection rates of antibody in camel sera. The IEP showed 84 p. 100 specificity and 36 p. 100 sensitivity while the IHA showed 69 p. 100 specificity and 43 p. 100 sensitivity. These results are discussed. 相似文献
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S. Sousa G. Thompson E. Silva L. Freire D. Lopes J. M. Correia da Costa A. Castro J. Carvalheira N. Canada 《Zoonoses and public health》2009,56(5):252-256
Toxoplasmosis is one of the most common food borne disease world‐wide. Among food animals, sheep seems to having higher prevalence of Toxoplasma gondii infection. However, there is no consensus about the best cut‐off for serodiagnosis in sheep. To estimate the more adequate cut‐off value of Modified Agglutination Test (MAT) for serodiagnosis in sheep, a commercial ELISA kit was used as a golden standard. Evaluation of the optimal sensitivity and specificity was calculated using Youden’s J‐statistics. Values obtained were used to estimate the prevalence of sheep toxoplasmosis. One thousand four hundred and sixty seven blood samples were collected randomly from 160 farms from northern Portugal, representing approximately 10% of the ovine population from the region. All sera were tested for anti‐T. gondii antibodies using the MAT. One hundred nine sheep (7.4%) presented a MAT titer ≥1 : 80; 45 (3.0%) a MAT titer of 1 : 40; 97 (6.6%) a MAT titer of 1 : 20 and 1216 (83.0%) a MAT titer ≤1 : 20. The best Youden’s J‐statistic was obtained at 1 : 20 titer (0.752), with 86.15% of sensitivity and 89.09% of specificity with negative and positive predictive values of 90.32% and 84.48% respectively, suggesting that the 1 : 20 was the most appropriate cut‐off for serodiagnosis of toxoplasmosis in sheep. Assuming this cut‐off, the prevalence of toxoplasmosis in the studied population was 17.1% and 92 (57.5%) of the 160 studied flocks having one or more positive sheep. Those results indicate that toxoplasmosis in Portugal should be considered in the differential diagnosis of abortions in sheep and neurological signs in lambs. Furthermore, while Portugal produces ovine meat for internal consumption and for exportation, isolation of T. gondii from ovine meat and further characterization of the isolates will be needed to understand the risk that ovine toxoplasmosis may represent for human health. 相似文献