采用不同的血清学检测法对羊布病检测的对比

本文采用虎红平板凝集试验、试管凝集试验和竞争ELISA试验对羊布病进行了对比检测,筛选出适合县级实验室对布病检测的最佳方法,为布病防控提供科学依据。     

布鲁氏菌病不同血清学检测方法结果分析

[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。     

山羊布鲁氏菌病不同血清学检测方法的比较

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。     

猪瘟的免疫血清学检测技术

本文综述了血清中和试验、猪瘟间接血凝试验、正向间接血凝试验、荧光抗体技术、免疫酶测定技术、间接ELISA、夹心-ELISA、DOT-ELISA、PPA-ELISA、C-ELISA、抗原捕获ELISA等在猪瘟血清学检测上的应用情况,并对其优、缺点作了浅析.     

布氏杆菌S2疫苗免疫绵羊血清抗体消长规律研究

为了解绵羊对布氏杆菌病S2疫苗免疫后的抗体消长规律,通过口服和注射2种方法对绵羊进行了布氏杆菌病S2疫苗免疫试验,利用虎红平板凝集试验(RBT)和试管凝集试验(SAT)以及c ELISA检测方法进行了血清抗体检测。结果表明:正常剂量口服免疫S2疫苗20 d后,RBT、SAT、c ELISA抗体阳性率均达到最高,分别是80%、70%、60%,并且在免疫100 d后羊血清抗体阳性率分别为RBT 20%、SAT 10%、c ELISA 10%,免疫250 d后3种方法检测结果均转为阴性。2倍剂量口服免疫S2疫苗20 d后,RBT、SAT、c ELISA均达到最高为60%,并在免疫100 d后均转为阴性。以正常剂量注射免疫S2疫苗免疫7 d后RBT和SAT抗体阳性率达到最高为80%,c ELISA抗体阳性率达到60%,30 d后达到80%,而疫苗免疫370 d后,RBT和SAT检测抗体阳性率下降到60%,c ELISA检测抗体阳性率依然80%;而2倍剂量注射免疫S2疫苗370 d后,RBT和c ELISA检测抗体阳性率仍保持着100%,SAT检测抗体阳性率达80%。结果表明,从免疫羊抗体产生及体内消长规律来看,布氏杆菌S2疫苗注射免疫明显优于口服免疫效果,且两种免疫途径的试验羊均且未发现明显接种副反应。     

不同布鲁氏菌血清学方法检测牛羊血清抗体的比对

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(cELISA)、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;cELI...     

对兽医实验室血清学抗体检测操作应用初探

近年来，基层（县级）兽医实验室相继通过省级兽医部门考核，已具备一级生物实验室相关要求，并逐步开展工作，但普遍存在化验操作不规范、化验结果不准确等现象。针对存在问题，结合几年来自身工作实践，现以猪瘟、口蹄疫血清学抗体检测过程为例，说明在实验操作过程中应注意的事项，供读者参考。     

Seroepidemiological investigation of brucellosis in sheep abortions in Kars,Turkey

This study was undertaken to investigate the seroprevalence of brucellosis in unvaccinated sheep from the flocks having previous abortion cases in Kars and around, Turkey and to compare the efficacy of each serological test used. Four hundred serum samples collected from 16 different flocks of sheep having a history of abortions in Kars and its surrounding area in Turkey were examined for the presence of antibodies raised against Brucella using Rose Bengal Plate Test (RBPT), Serum Agglutination Test (SAT), Rivanol Agglutination Test (RAT) and Complement Fixation Test (CFT). All animals were unvaccinated against Brucella. Of the serum samples tested, 147 (%36.7), 142 (%35.5), 139 (%34.75) and 135 (%33.75) were found positive by SAT, RAT, RBPT and CFT, respectively. No statistically significant difference was found between the serological tests used (p > 0.05). It is concluded from this study that brucellosis continues to be an important problem for ovine abortions and poses a risk both for human and other animals in this area. Therefore, adequate intervention measures should be implemented to control and eradicate brucellosis. In addition, if conventional serological tests are used at least two tests, RPBT for screening and CFT for the confirmation of the positive samples are preferable, should be used in parallel for detection of brucellosis effectively. This study was summarized from M.Sc. thesis of Ozgur CELEBI.     

布鲁氏菌弱毒疫苗粘膜免疫及检测方法的研究

为研究布鲁氏菌弱毒疫苗粘膜免疫及其检测方法,本实验采用粘膜点眼途径对健康母羊接种布鲁氏杆菌猪2号疫苗(S2)、牛19号疫苗(A19)和羊强毒株(M16),筛选布鲁氏杆菌病鉴别检测方法。将12月龄～14月龄母羊60只随机分为3组,以常规疫苗推荐剂量进行半量粘膜点眼接种。采集血液、淋巴、脏器进行布鲁氏菌病血清学检测和细菌学分离以及PCR检测。结果表明:布鲁氏菌弱毒疫苗抗体水平持续6个月,其中血清学的试管凝集试验、半胱氨酸凝集试验与补体结合试验的阳性符合率达到100%。细菌分离期为6个月,乳腺、乳腺淋巴、髂淋巴分离率较高;而强毒株M16的抗体水平和细菌分离持续12个月以上。结果显示以常规血清学和细菌学检测方法在点眼免疫布鲁氏菌S2、A19苗6个月后可以进行野毒感染和疫苗免疫畜的鉴别诊断。     

Assessing PCV2 antibodies in field pigs vaccinated with different porcine circovirus 2 vaccines using two commercial ELISA systems

Porcine circovirus type 2 (PCV2) is the primary causative agent for post-weaning, multisystemic, wasting syndrome. Consequently, serologic detection of and vaccination against PCV2 are important for the swine industry. Among several serological tests, the enzyme-linked immunosorbent assay (ELISA) is commonly used to measure anti-PCV2 antibody levels. In the present study, we used two commercial ELISA systems to comparatively evaluate anti-PCV2 antibodies in field pigs treated with three different PCV2 vaccines. Among a total of 517 serum samples, the results of the two ELISAs were fully concordant for 365 positive and 42 negative samples, indicating 78.7% agreement. In addition, the Pearson coefficient (0.636) indicated a moderate correlation between data from the two ELISAs. Results from the farms with pigs vaccinated with the three different PCV2 vaccines demonstrated that most of the vaccinated animals underwent seroconversion. However, the increase and duration of antibody titers varied depending on the vaccine, the presence of maternal antibodies, and the vaccination program. PCV2 serologic status and anti-PCV2 antibody levels of herds from this study could be utilized to determine the best timing for vaccination and assessing vaccination compliance.     

Agglutination tests and their modifications in the diagnosis of bovine brucellosis

This article reviews the effects of using Yersinia enterocolitica serotype: 09 somatic antigen in diagnostic tests for brucellosis. The tests employed include the standard tube agglutination test, the microplate agglutination test, the quantitative plate agglutination test, the growth agglutination test and the indirect hemagglutination technique. Only the growth agglutination technique demonstrated an increased sensitivity over the STAT. Neither the microplate agglutination test nor quantitative plate tests offered any significant advantages over the STAT. The QPAT using Brucella O antigen was clearly subject to false negative results. The indirect hemagglutination technique showed extreme variation in results, the significance of which (if any) was unclear.     

Assessment of performance of selected serological tests for diagnosing brucellosis in pigs

Swine brucellosis due to Brucella suis is considered an emerging zoonotic disease whose control is based on serological testing and the subsequent culling of seropositive animals or the full depopulation of affected flocks. Here we assessed the performance of several serological tests (Rose Bengal Test [RBT], indirect ELISA [i-ELISA], blocking ELISA [b-ELISA], and two competitive ELISAs [c-ELISA]) for diagnosing swine brucellosis caused by B. suis biovar 2. Both frequentistic and Bayesian statistical inference were used. A frequentistic analysis, using sera from known gold standard (GS) populations (i.e., from truly infected or brucellosis free animals), resulted in maximum (100%) diagnostic sensitivity (Se) and specificity (Sp) in the RBT, i-ELISA and b-ELISA tests. However, c-ELISAs resulted in lower diagnostic Se (ranging from 68.5% to 92.6%, according to the different cut-offs selected). A Bayesian analysis of tests yielding the best diagnostic performance with GS sera (RBT, i-ELISA and b-ELISA), but using a large collection of field sera, resulted in similar Se among tests but markedly lower (≈ 80%) than that resulting from the frequentistic analysis using the GS serum populations. By contrast, the estimated Sp in the Bayesian analysis was only slightly lower than 100%, thus similar to that obtained frequentistically. Our results show that adequate diagnostic tests for brucellosis in swine are available, but also emphasize the need for more extensive validation studies before applying these tests under field conditions.