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1.
旨在科学选择和使用布鲁氏菌抗体检测方法,推动布病诊断试剂标准化。本研究用布病阳性血清标准品测定了国家/OIE布鲁氏菌病参考实验室开发的布鲁氏菌荧光偏振(FPA)抗体检测试剂盒、动物布鲁氏菌病竞争ELSIA (cELISA)抗体检测试剂盒、牛布鲁氏菌病间接ELISA (iELISA)抗体检测试剂盒和改进的微量补体结合试验(mCFT)等4种方法的灵敏度。通过对已知阴、阳性血清样品的检测,比较了各检测方法的敏感性和特异性,并用临床样本进一步比较了各种方法检测结果的吻合性。结果表明,4种方法检测的灵敏度基本一致,当布病阳性血清标准品按1∶20稀释(即50 IU·mL-1)时均检测为阳性,1∶40稀释(即25 IU·mL-1)时均检测为阴性。FPA、cELISA、iELISA和mCFT方法的敏感性分别为97.14%、100.00%、100.00%、98.57%,特异性分别为96.34%、95.12%、97.56%、100.00%。对315份临床样本的检测结果显示,各方法之间的符合率均高于90.00%,其中iELISA、FPA、cELISA与mCFT符合率分别为97.14%、96.83%、92.70%;FPA、cELISA与iELISA符合率分别为95.24%、93.65%;FPA与cELISA符合率为91.43%。iELISA、FPA、mCFT 3种方法之间吻合性最高,cELISA与其他3种方法之间的吻合性略低。  相似文献   

2.
为评价不同布鲁氏菌血清学方法检测牛羊血清的性能,采用试管凝集试验(SAT)、竞争酶联免疫吸附试验(c ELISA)、荧光偏振试验(FPA)3种常规血清学检测方法,对已知感染情况的牛羊血清进行布鲁氏菌抗体检测,比较这3种方法的一致性、敏感性与特异性。结果显示:SAT检测牛血清的Kappa值小于0.75,一致性一般;cELISA、FPA检测牛羊血清的Kappa值均大于0.75,一致性较好。SAT特异性较高,假阳性率均为0,但敏感性较低,假阴性率较高;cELISA敏感性和特异性较为理想,均在92%以上;FPA敏感性最好,为100%,特异性也较高,达97%以上。结果表明,SAT特异性高,但敏感性略低,c ELISA和FPA敏感性、一致性和特异性均较好。结果提示,在实际诊断中,先用操作简便的SAT进行筛选,再用操作较为复杂、需要特定仪器、特异性高的cELISA和FPA进行复核较为理想。  相似文献   

3.
《畜牧与兽医》2016,(7):113-114
旨在比较奶牛布氏杆菌病3种血清学检测方法,为临床根据实际检测情况选择不同检测方法提供参考。对420头奶牛血清样品用虎红平板凝集试验(RBT)、试管凝集试验(SAT)、酶联免疫吸附试验(ELISA)3种方法进行布病抗体检测,比较总符合率、假阴性、阳性率及敏感性和特异性。结果:RBT、SAT、ELISA 3种检测方法的总符合率高达到90%以上;以SAT作为判定标准,ELISA检测结果的假阴性率(7.1%)和假阳性率(1.3%)均低于RBT检测结果的假阴性率(14.3%)和假阳性率(3.1%),而ELISA的敏感性(92.3%)和特异性(98.7%)均高于RBT的敏感性(85.7%)和特异性(96.9%)。临床工作中使用RBT或者ELISA进行初步筛选,SAT进行复核确诊,2种或2种以上的检测方法联合运用结果较为理想。  相似文献   

4.
为比较山羊布鲁氏菌病不同血清学检测方法,继而为山羊布鲁氏菌病的临床检测提供参考,对采集到的423份血清样品分别用虎红平板凝集试验(RBT)、试管凝集试验(SAT)和竞争酶联免疫吸附试验(c ELISA)3种方法进行检测,比较3种方法的一致性、敏感性和特异性。结果表明:RBT和SAT、cELISA和RBT、c ELISA和SAT的Kappa值均大于0.75,一致性较好;RBT和SAT符合率最高,RBT敏感性最好,cELISA特异性最好。因而建议在开展山羊布鲁氏菌病检测时,可先用RBT初筛,再用SAT或c ELISA复检。  相似文献   

5.
[目的 ]为在布鲁氏菌病临床检疫中选择可靠的血清学检测方法提供参考。[方法 ]对采集的294份牛血清样品用虎红平板凝集试验(RBT)、试管凝集试验(SAT)、酶联免疫吸附试验(ELISA)和补体结合试验(CFT)进行布鲁氏菌病抗体检测,比较RBT与ELISA,SAT与CFT的符合率及Kappa值,以CFT作为判定标准,比较四种检测方法的敏感性和特异性。[结果 ]RBT与ELISA、SAT与CFT检测方法的符合率高达到95%以上,且Kappa值均大于0.75。以CFT作为判定标准,RBT和ELISA的敏感性较好,但有假阳性;SAT的特异性较好,但有假阴性。综合比较认为ELISA的敏感性和特异性都比较理想。[结论 ]临床工作中使用RBT或ELISA对布鲁氏菌病进行初筛,用CFT进行复核确诊,通过两种或两种以上的血清学检测方法联合诊断,结果较为理想。  相似文献   

6.
为选择科学适用的牛布鲁氏菌病净化检测方法,对1 721份免疫布鲁氏菌A19号疫苗18个月后的牛血清,分别用虎红平板凝集试验(RBT)、试管凝集试验(SAT)、竞争酶联免疫吸附试验(cELISA)、间接酶联免疫吸附试验(iELISA)和荧光偏振试验(FPA)5种血清学检测方法,以及环介导等温扩增技术(LAMP)和荧光定量 PCR病原学检测方法进行检测,分析比对不同检测方法的特异性、敏感性、误诊率、漏诊率、符合率、Kappa值等。结果显示:与SAT相比,其他4种血清学检测方法的敏感性、特异性和符合率从高到低依次为iELISA、FPA、cELISA、RBT,其中敏感性均在85.00%以上,特异性均在97.60%以上,符合率均在97.50%以上,Kappa值均≥1.00;与PCR相比,LAMP方法敏感性低(9.09%),但特异性强(99.65%),与PCR符合率高(98.49%)。结果表明:血清学检测方法较为敏感特异,符合率和一致性均较高,而分子生物学检测方法特异性强,不易误诊和漏诊。因此,对于牛群的布鲁氏菌病净化,要结合牛群污染和免疫情况以及净化的不同阶段选择适用的检测方法,仅用一种方法可能会存在偏差。建议先用iELISA或FPA或RBT进行初筛,再用cELISA或SAT进行确诊,进而对确诊为阳性牛的阴道拭子或奶样进行分子生物学检测,以确定是否存在布鲁氏菌感染。本研究为免疫牛群的布鲁氏菌病净化检测方法选择提供了参考。  相似文献   

7.
《中国兽医学报》2020,(2):336-338
为了比较国产布鲁菌病间接ELISA抗体检测试剂盒、竞争ELISA抗体检测试剂盒和胶体金试纸条的检测效果,通过对已知阴阳性血清样品的检测,比较了上述3种检测方法的敏感性和特异性。进一步采用临床血清样品比较了3种检测方法与虎红平板凝集试验(RBT)的检测效果,并采用补体结合试验(CFT)对结果进行了复核。结果显示,间接ELISA、竞争ELISA和胶体金试纸条的敏感性分别为96.67%,100.00%和98.33%,3种检测方法的特异性分别为98.33%,93.33%和93.33%。对300份临床样品的检测结果显示,4种检测方法共同检出的阳性样本为27份,阴性样品为210份,整体符合率为79%。通过CFT对63份不同方法检验结果有差异的样本进行确诊,结果表明RBT与CFT的符合率最低,仅为3.17%;间接ELISA与CFT的符合率最高,为98.41%;竞争ELISA和胶体金试纸条的符合率分别为87.30%,85.71%。结果表明,间接ELISA、竞争ELISA和胶体金试纸条具有良好的特异性和敏感性,能够满足布病临床检测需求;RBT对临床样本的检测存在较高比例假阳性和假阴性。  相似文献   

8.
[目的]探究湖羊经布鲁氏菌S2疫苗免疫后的血清抗体消长规律。[方法 ]采用皮下注射25亿CFU(正常剂量)、口服免疫100亿CFU(正常剂量)、口服免疫150亿CFU(1.5倍剂量)3种免疫方法,对试验湖羊进行S2疫苗接种。在免疫前后的不同时间点,采集血液,分离血清,通过虎红平板凝集试验(RBT)和试管凝集试验(SAT)检测血清抗体水平。[结果 ]皮下免疫组在免疫后7 d出现抗体,21 d达到峰值,平均抗体效价最高为1:400,RBT和SAT抗体阳性率均为100%;免疫后120 d,RBT、SAT血清抗体阳性率分别为53%、56%;血清抗体存续时间为240 d。1.5倍剂量口服组免疫后21 d,RBT、SAT抗体阳性率分别为80%、83%,平均抗体效价最高为1:230.9,血清抗体存续时间为150 d。正常剂量口服免疫组在免疫后14 d产生抗体,21 d的抗体阳性率最高,RBT、SAT抗体阳性率分别为50%、57%,平均抗体效价最高为1:156,血清抗体存续时间为120 d。[结论 ]布鲁氏菌S2疫苗皮下免疫效果明显优于口服免疫效果,但可能存在免疫副反应;1.5倍剂量口服优于正常剂量口服。  相似文献   

9.
为比较牛羊布鲁氏菌竞争酶联免疫吸附试验(cELISA)与其他免疫学方法检测的一致性,使用2种国产布鲁氏菌cELISA试剂盒以及虎红平板凝集试验(RBT)、试管凝集试验(SAT)方法,对采集自山东省日照市的122份牛羊血清样品分别开展布鲁氏菌抗体检测,比较2种cELISA试剂盒与RBT、SAT检测符合率和Kappa值。结果显示,2种cELISA试剂盒与RBT、SAT的符合率均在82%以上,Kappa值均大于0.61。结果表明,2种国产cELISA试剂盒适于牛羊布鲁氏菌净化检测。本文为牛羊布鲁氏菌血清学检测方法的选择应用提供了参考。  相似文献   

10.
荧光偏振试验(FPA)是一种新的试验方法。本文应用荧光偏振检测方法对出口哈萨克斯坦的321份牛进行布氏杆菌病检测,结果有25份牛血清样品检测为抗体阳性。同时对321份血清进行虎红平板凝集试验(RBPT)、试管凝集试验(SAT)、补体结合试验(CFT)、间接ELISA试验(I-ELISA)、竞争ELISA试验(C-ELISA)等比对试验,结果是FPA从敏感性和特异性上都优于或等于目前主要采用的RBPT、SAT、CFT、ELISA检测方法。FPA方法简便、快速、通量大,不需洗板,可在15分钟内完成92份样品的布氏杆菌病检测,极适合于大批量牛布氏杆菌病的检疫、筛查和疫病监控。  相似文献   

11.
Serological methods are traditionally used in diagnosis of brucellosis. However, the comparative performance of these tests and their accuracy under the local environment in Zambia has not been assessed. Thus, the objective of our study was to evaluate the diagnostic performance of three serological tests for brucellosis; Rose Bengal Test (RBT), competitive ELISA (c-ELISA) and Fluorescence Polarisation Assay (FPA) in naturally infected cattle in Zambia without an appropriate reference test to classify animals into truly infected and non-infected. Serological test results from a study to determine sero-prevalence were used to compare the performance of RBT, c-ELISA and FPA in diagnosing brucellosis in traditional cattle. Since none of the tests can be seen as a perfect reference test or gold standard, their performance in a population of naturally infected cattle was evaluated using latent class analysis which allows the sensitivity (Se) and specificity (Sp) to be estimated in the absence of a gold standard. The highest Se was achieved by the c-ELISA (97%; Credible Posterior Interval (CPI)=93-100%) and the highest Sp by the FPA (93%; CPI=85-99%), conversely these tests also had the lowest Sp and Se, respectively, with the RBT performing well in both the Se (93%; CPI=84-98%) and Sp (81%; CPI=61-97).  相似文献   

12.
The serological response of young and adult sheep vaccinated conjunctivally with Rev-1 vaccine was assessed by fluorescence polarization assay (FPA), Rose Bengal test (RBT), complement fixation test (CFT), modified Rose Bengal test (m-RBT), indirect ELISA (i-ELISA) and competitive ELISA (c-ELISA), at different post vaccination intervals. One hundred and thirty six adult sheep and 64 lambs were used in the study. The vaccinated animals were bled prior to vaccination (0 day) and thereafter at 21st, 42nd, 35th, 63rd, 91st, 125th, 159th, and 223rd and 330th day post vaccination. The majority of animals (young and adult) showed positive reaction by FPA, RBT, CFT, m-RBT and c-ELISA 21 days post vaccination, whereas by i-ELISA at 42 days. All tests perform equal when animals vaccinated as young are tested 125 days (4 months) post vaccination. In case of animals vaccinated at adulthood, FPA, RBT, CFT and c-ELISA perform equal if the animals are tested 223 days (approximately 8 months) post vaccination. I-ELISA and m-RBT show low specificity if ewes vaccinated at adulthood are tested 330 days (11 months) post vaccination. If control of brucellosis in sheep is based on conjunctivally vaccination of lambs with Rev-1, the vaccinated animals can be tested by any test used for diagnosis of B.melitensis infection accurately at least 4 months post vaccination. If brucellosis control is based on mass vaccination the use of m-RBT and i-ELISA is not recommended for testing adult animals at least for 330 days (11 months) post vaccination due to tests low specificity. Further research is needed so the appropriate cut-offs to be established for FPA, c-ELISA or i-ELISA to become valuable tools for the eradication of Brucella spp. infection in small ruminants in areas where vaccination is practiced.  相似文献   

13.
The effectiveness of Rose Bengal test (RBT) and fluorescence polarization assay (FPA) in diagnosing cattle brucellosis in endemic areas was assessed and RBT and FPA test agreement was compared (n = 319). The sensitivity of RBT and FPA in detecting low Brucella titres were evaluated in paired sera (n = 34). A logistic regression model was constructed to predict cattle test result in FPA using RBT as the main predictor and incorporating bio-data and animal history. There was 79.3% agreement between the RBT and FPA (Kappa = 0.59; Std error = 0.05; p = 0.000) and a high correspondence between high RBT scores and positive FPA results suggesting that sera with high RBT score may not require confirmation with tests such as competitive-ELISA or CFT. High FPA cut-off points were more likely to miss animals with low antibody titres. The RBT had a reduced ability in detecting low antibody titres compared to the FPA. FPA test interpretation was improved if a priori information, such as sex and age was used. Under the challenging disease surveillance conditions prevailing in rural Africa, field-testing methods that are sensitive and specific; allow single animal contact, low technical skills in data interpretation are suitable.  相似文献   

14.
The fluorescence polarization assay (FPA) was evaluated for the serological diagnosis of brucellosis in water buffalo (Bubalus bubalis) in southern Italy. This assay uses O-polysaccharide prepared from Brucella abortus lipopolysaccharide conjugated with fluorescein isothiocyanate as a tracer. It has many methodological advantages over older, more established tests and can be performed in a fraction of the time. Sera from 890 buffalos from the Campania Region - 526 positive sera and 364 negative sera according to the complement fixation test (CFT) - were evaluated in this study. All samples were tested with the Rose Bengal test (RBT), CFT, and FPA in parallel and in blind fashion. Sensitivities (Sn) were 84.5% and 92.6%, and specificities (Sp) were 93.1% and 91.2% for RBT and FPA, respectively, relative to CFT. Finally, receiver operating characteristic (ROC) analysis suggested a cut-off value of 117 millipolarization (mP) units. On the whole, these results suggested that FPA might replace RBT in the diagnosis of buffalo brucellosis for its better performance relative to CFT, its adjustable cut-off useful in different epidemiological situations, its reliability, ease of performance, and for its potential application in field and high-throughput laboratories.  相似文献   

15.
An automated indirect enzyme-linked immunosorbent assay (I-ELISA) for the serological diagnosis of bovine brucellosis was developed and validated in-house. A total of 4,803 cattle sera from South Africa (n = 3,643), Canada (n = 652), Germany (n = 240), France (n = 73) and the USA (n = 195) was used. The South African panel of sera represented 834 sera known to be positive by the Rose Bengal test (RBT), serum agglutination test (SAT) and complement fixation test (CFT), 2709 sera that were negative by CFT, and 100 sera from animals vaccinated with a standard dose of Brucella abortus strain 19. Overseas sera were obtained from reference non-vaccinated brucella-free cattle (n = 834), naturally infected (n = 72), experimentally infected (n = 71), and vaccinated animals (n = 83). Also 100 sera collected from cattle in Canada and known to be positive by competitive ELISA (C-ELISA) were used. The intermediate ranges ("borderline" range for the interpretation of test results) were derived from two-graph receiver operating characteristics analysis. The lowest values of the misclassification cost-term analysis obtained from testing overseas panels, covered lower I-ELISA cut-off PP values (0.02-3.0) than those from local panels (1.5-5.0). The relatively low cut-off PP values selected for I-ELISA were due to the fact that the positive control used represents a very strong standard compared to other reference positive sera. The greater overlap found between negative and positive cattle sera from South Africa than that between reference overseas panels was probably due to the different criteria used in classifying these panels as negative (sera from true non-diseased/non-infected animals) or positive (sera from true diseased/infected animals). The diagnostic sensitivity of the I-ELISA (at the optimum cut-off value) was 100% and of the CFT 83.3%. The diagnostic specificity of I-ELISA was 99.8% and of the CFT 100%. Estimate of Youden's index was higher for the I-ELISA (0.998) than that for the CFT (0.833). Analysis of distribution of PP values in sera from vaccinated and naturally infected cattle shows that in vaccinated animals all readings were below 31 PP where in infected ones these values represented 43%. Therefore, it appears that I-ELISA could be of use in identifying some naturally infected animals (with values > 31 PP), but more sera from reference vaccinated and infected animals need to be tested to further substantiate this statistically. Of 834 sera positive by RBT, SAT and CFT, 825 (98.9%) were positive in the I-ELISA. Compared to C-ELISA the relative diagnostic sensitivity of the I-ELISA was 94% and of the CFT 88% when testing 100 Canadian cattle sera. Of 258 South African cattle sera, of which 183 (70.9 %) were positive by the I-ELISA and 148 (57.4 %) by the CFT, 197 (76.4%) were positive by C-ELISA when re-tested in Canada. One has to stress, however, that Canadian C-ELISA has not been optimised locally. Thus, the C-ELISA was probably not used at the best diagnostic threshold for testing South African cattle sera. This study shows that the I-ELISA performed on an automated ELISA workstation provides a rapid, simple, highly sensitive and specific diagnostic system for large-scale detection of antibodies against B. abortus. Based on the diagnostic accuracy of this assay reported here, the authors suggest that it could replace not only the currently used confirmatory CFT test, but also the two in-use screening tests, namely the RBT and SAT.  相似文献   

16.
While serological tests are essential in surveillance and control programs of animal diseases, to date none of the common serological tests approved in the EU (complement fixation test or Rose-Bengal test) has been shown to be reliable in routine individual diagnosis of porcine brucellosis, and some more recent tests like ELISA have not been fully evaluated yet. In the absence of a gold standard, this study allowed the estimation of sensitivities and specificities of these tests with a Bayesian approach using Markov Chain Monte Carlo algorithms. The pig population that was tested included 6422 animals from Metropolitan France. Serum samples were collected from a large population of pigs, representative of European swine population and tested with five brucellosis serological tests: Rose-Bengal test (RBT), fluorescence polarization assay (FPA), indirect ELISA (I-ELISA) and two competitive ELISAs (C-ELISA). The sensitivity and the specificity of each test were estimated. When doubtful results were excluded, the most sensitive and specific test was C-ELISA(2) (Se C-ELISA(2)=0.964, [0.907; 0.994], 95% credibility interval (CrI); Sp C-ELISA(2)=0.996, [0.982; 1.0], 95% CrI). When doubtful results were considered as negative, C-ELISA(2) was still the most sensitive and specific test (Se C-ELISA(2)=0.960, [0.896; 0.994], 95% CrI and Sp C-ELISA(2)=0.994, [0.977; 0.999], 95% CrI). The same conclusions were reached when doubtful results were considered as positive (Se C-ELISA(2)=0.963, [0.904, 0.994], 95% CrI and Sp C-ELISA(2)=0.996, [0.986; 1.0], 95% CrI).  相似文献   

17.
Three serological methods, the Rose-Bengal test (RBT), the complement-fixation test (CFT) and an indirect enzyme-linked immunosorbent assay (I-ELISA) were compared for the detection of Brucella-infected animals in unvaccinated cattle herds in Eritrea. In this study, 71 herds first were classified as positive or negative for Brucella infection on the basis of at least one animal being seropositive by RBT and CFT. All the 159 RBT-positive samples from the 26 seropositive herds and 214 RBT-negative samples randomly selected from the seropositive herds and from the 45 negative herds were tested further by CFT and I-ELISA. Using the ELISA titer as main predictor, and incorporating the RBT results, a logistic model was built to predict the CFT-negative or -positive status of individual sera and to estimate sensitivity and specificity. Whilst the ELISA titers (< or =20) accurately predicted all the negative sera in herds that were also negative by the CFT, the number of seropositive animals was higher by ELISA in herds that had positive animals. Serum samples which give higher degrees of agglutination with the RBT need not be re-tested with CFT; consideration of the seropositive status of a herd should be taken into consideration on defining the cut-off optical density readings for ELISA.  相似文献   

18.
An evaluation of fluorescence polarization assay (FPA) to detect antibodies against Brucella melitensis according to the Mexican Official Norm (NOM) was performed. In this study, a total of 2582 goat serum samples from a high-prevalence area in northeast Mexico where vaccination is applied, were used. Of these, 1094 were classified as NOM negatives (card test (CT) negatives or CT positives/complement fixation test (CFT) negatives) and 1488 as NOM positives (CT and CFT positives). The receiver operator characteristics (ROC) curve analysis was used to obtain the FPA sensitivity (83.5%), specificity (82.2%) and accuracy (88.2%) compared with NOM criteria, using a cut-off value of 89mP for positive samples. In addition, FPA produced 84.1% of negative results versus 65.7% of CT using 1094 CFT negative samples, which indicated that FPA performance was better than CT to detect negative samples or differentiate samples from vaccinated animals. Finally, FPA showed 95.8% sensitivity when using 702 negative non-vaccinated samples. Taken together, these results suggested that FPA might replace CT as a screening test for its better performance compared with CFT, its adjustable cut-off useful in different epidemiological situations, and for its reliability, ease of performance, comparable cost with CT regimen, and potential application in field and high-throughput laboratories. The use of FPA as screening test will help to reduce the percentage of goats wrongly slaughtered because of brucellosis misdiagnosis. More studies on FPA are required for its approval as diagnostic tool for goat brucellosis.  相似文献   

19.
布鲁菌病主要采用凝集试验来检测,本试验用虎红平板凝集试验和试管凝集试验对江苏海安、江都、无锡和兴化地区的660头奶牛的血清样品进行平行检测,虎红平板凝集试验的阳性检出率为9.7%,试验管凝集试验的阳性检出率为5.5%;与后者相比,虎红平板凝集试验的敏感性为100%,特异性为95.3%,符合率为95.6%。这些数据表明,江苏地区某些奶牛场存在布鲁菌感染,虎红平板凝集试验可以用于布鲁菌病初步检测。  相似文献   

20.
[目的] 为了确定不同布病检测方法在临床应用中的意义。[方法] 对疑似感染布鲁氏菌病羊血清同时作虎红平板凝集实验(RBT)、试管凝集实验(SAT)、ELISA抗体检测(cELISA)检测,分析它们的Kappa值、符合率、敏感度和特异性等。[结果] 数据显示三者符合度较高,Kappa值在0.76以上,一致性相当可靠,其中RBT敏感度高,但假阳性率也高,相比SAT结果,cELISA敏感度和特异性都高,且两者Kappa值达到了0.85,综合判断临床工作中cELISA更适合确诊诊断。  相似文献   

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