布鲁氏菌病三种诊断方法的比较

采集太原市周边县区4个规模场的牛血清样品400份,用虎红平板凝集试验（RBPT）、试管凝集试验（SAT）、iELISA进行抗体检测,比较三种检测方法的敏感性和特异性差异。结果表明三种方法检测结果一致率较高,但是RBPT和SAT均有较高的假阳性和假阴性,三种方法中iELISA的敏感性和特异性最好。结论是在布病检测时可选择操作简便的RBPT进行初筛,阳性结果用iELISA进行确诊。     

几种布鲁氏菌病检测方法比对结果分析

为比较几种常用布鲁氏菌病(简称布病)检测方法特性,在某疫苗免疫的奶牛场随机选取40头奶牛,采集血清和奶样各40份,然后对采集的血清进行虎红平板凝集、试管凝集、胶体金、酶联免疫吸附试验(iELISA和cELISA)和琼脂扩散方法检测,对采集的奶样进行病原学检测(qPCR)和抗体检测(胶体金)。结果显示:4种国标方法(虎红平板凝集、试管凝集、iELISA和cELISA)检出的阳性样品数量有较大差距,分别为16、6、23和33份,其中经试管凝集试验检出的阳性血清,经其他3种国标方法也检测为阳性;通过胶体金法检出阳性血清数(9份)略高于试管凝集试验,且血清及牛奶样品经胶体金检测结果基本一致;琼扩法检出12份阳性血清,且显示抽检奶牛中存在野毒感染;qPCR和胶体金试纸条对采集奶样的检测结果有较大差距,经qPCR检测仅1份样品呈阳性。结果表明:国标方法中,试管凝集试验特异性最高,iELISA和cELISA敏感性较高;病原学检测临床样品检出率较低,琼脂扩散试验敏感性有限,胶体金法敏感性适中,操作简便,适合布病免疫奶牛场自检。本研究为牛场布病不同检测目的方法选择提供了参考。     

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。     

牛布鲁氏菌A19号疫苗免疫血清几种检测方法的比较

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。     

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。     

奶牛布氏杆菌病3种血清学检测方法的临床应用比较

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。     

牛羊布鲁氏菌cELISA与RBT、SAT检测方法比较

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。     

几种布鲁氏菌病检测方法的比较

为研究虎红平板凝集试验(RBT)、全乳环状试验(MRT)、间接酶联免疫吸附试验(iELISA)及胶体金检测试纸条对布病的检测效果,本研究采用以上方法分别对感染地区和净化地区的牛群血清、羊群血清、牛群奶样进行了检测效果的比对。结果显示,针对牛群血清,RBT结果阳性率最高,iELISA与胶体金结果阳性率相似,表明RBT的敏感性较好,iELISA与胶体金检测试纸条的特异性较RBT好;针对羊群血清,iELISA与胶体金检测试纸条的敏感性和特异性较RBT好;针对牛群奶样,MRT结果阳性率低于全乳iELISA和胶体金检测试纸条。全乳iELISA和胶体金检测试纸条在试验中显示出具有较高的敏感性和特异性。     

牛羊布鲁氏菌病四种血清学检测方法对比分析

[目的]对布鲁氏菌病的几种检测方法进行比对分析,为国家标准修订提供参考。[方法]对收集到的669份血清样本用虎红平板凝集试验（RBT）、试管凝集试验（SAT）、补体结合试验（CFT）和酶联免疫吸附试验（ELISA）方法进行检测,比较四种检测方法的一致性、敏感性与特异性。[结果]RBT、SAT、CFT、ELISA四种牛羊布鲁菌病的血清学检测方法一致率较高,Kappa值均大于或等于0.75。RBT敏感性较高,CFT特异性好,而SAT有一定的假阳性和假阴性,ELISA的敏感性和特异性都比较理想。[结论]用RBT初筛,用CFT和ELISA联合诊断结果较为理想。     

牛布鲁氏菌四种血清学检测方法的比较

[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。     

Field trial of six serological tests for bovine brucellosis

Serum agglutination (SAT), complement fixation (CFT), indirect ELISA (iELISA), competitive ELISA (cELISA), Rose Bengal (RBT) and EDTA-modified agglutination (EDTA) tests were used in parallel on serological samples from 19,935 cattle in 301 herds. The study herds were selected according to putative exposure to Brucellaabortus with cases defined by bacteriological culture or test agreement. No single test identified all infected cattle and, at diagnostic thresholds, relative sensitivity was highest in the iELISA (67.9%) or RBT (78.1%), using bacteriological culture or test agreement, respectively, to define cases. As screening tests, the relative sensitivity of the SAT was highest (75.9% by culture or 84.9% by test agreement), with an optimal threshold of 31 IU. The relative specificity of the diagnostic tests ranged from 99.6% (SAT 31IU) to 100% (iELISA, RBT and CFT). The trial confirmed the value of the SAT as a screening test and the value of parallel testing.     

布鲁氏菌4种高通量抗体检测方法的比较研究

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。     

虎红平板凝集试验与试管凝集试验检测奶牛布鲁菌病的比较

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。     

Assessment of milk ring test and some serological tests in the detection of <Emphasis Type="Italic">Brucella melitensis</Emphasis> in Syrian female sheep

Brucella melitensis infection prevalence among Syrian female sheep, to evaluate a number of serological tests and to discuss some epidemiological aspects of brucellosis, was studied. A total of 2,580 unvaccinated Syrian female sheep sera samples were tested for B. melitensis antibodies detection using four serological methods: the Rose Bengal test (RBT), the serum agglutination test (SAT), the complement fixation test (CFT) and an indirect enzyme-linked immunosorbent assay (iELISA). In addition, 2,375 milk samples were collected, then milk ring test (MRT) and bacterial isolation test were employed to evaluate the natural organism shedding. The samples were considered positive in 66%, 64%, and 60% when we employed the RBT, SAT, and iELISA tests, respectively. Whereas, the CFT test revealed the smallest number of positive samples. By using the MRT, the total prevalence of brucellosis was nearly 38% of samples. A large variation was observed concerning the studied areas, ranging from 24% in Tartous to 44% in both Damascus and Damascus rural areas. Brucella was isolated from only 677 samples out of the 2,375 female sheep milk samples.     

补体结合酶联免疫吸附试验方法的建立

为改进免疫学诊断技术的准确性,研究了一种基于补体结合的免疫学检测新技术———补体结合酶联免疫吸附试验（CF-ELISA）。CF-ELISA技术采用酶标记抗菊糖纯化豚鼠补体C3抗体及其酶显色系统作为补体参与反应的指示系统,用ELISA方法进行补体结合试验。经对布氏菌病抗体检测的初步试验结果显示,CF-ELISA技术可检测到0.01 IU的布氏菌病抗体,灵敏度与间接酶联免疫吸附试验（iELISA）相当,是虎红平板凝集试验（RBPT）试管凝集试验（SAT）的5 000倍、补体结合试验（CFT）的10 000倍。对349份确诊布氏菌病感染群牛、羊血清的检测结果显示,CF-ELISAi、ELISA、CFT、SAT、RBPT的阳性率分别为35.82%3、6.39%、31.81%、30.09%、36.1%,CF-ELISA与iELISA、CFT、SAT、RBPT的阳性符合率分别为：98.4%、88.8%、80.0%、90.6%。CF-ELISAi、ELISA、CFT、SAT、RBPT对490份布氏菌病阴性群牛、羊血清的阴性率分别为100%、99.6%、100%、99.4%、99.8%,CF-ELISA与iELISA、CFT、SAT、RBPT的阴性符合率分别为：99.6%1、00%、99.4%、99.8%。研究表明,CF-ELISA是具有高特异性和高敏感性的布氏菌病免疫学检测技术。     

不同方法检测奶牛布氏杆菌抗体的比较试验

采用虎红平板凝集试验(RBT)、全乳环状试验(MRT)和试管凝集试验(SAT)3种国家标准认可的动物布病检疫方法检测奶牛布氏杆菌抗体,并进行了比较研究。从82头非免疫和S2菌苗免疫的奶牛采集血样和乳样,RBT检测血样的阳性率为51.22%(42/82),MRT检测乳样的阳性率为46.34%(38/82),SAT检测血样的阳性率为45.12%(37/82)。以SAT定量检测的结果为参照标准,RBT初筛试验的符合率比MRT高,而且MRT反应强度的差异与SAT抗体滴度没有明显相关性。另对乳清和血清样本中布氏杆菌抗体的差异进行了分析讨论。     

Bayesian estimation of the true prevalence,sensitivity and specificity of the Rose Bengal and indirect ELISA tests in the diagnosis of bovine brucellosis

Serology is the most convenient method for detecting brucellosis but the efficient use of such tests in disease control requires evaluation of diagnostic performance and discriminative ability. The objective of this study was to assess the performance of the Rose Bengal test (RBT) and an indirect ELISA (iELISA) in diagnosing brucellosis in 995 serum samples collected from cattle in the Ivory Coast between 2005 and 2009. A Bayesian approach was used to evaluate the two tests by estimating their sensitivities and specificities.The correlation-adjusted sensitivity of the iELISA was estimated to be 96.1% (credibility interval [CrI], 92.7–99.8), whereas that of the RBT was 54.9% (CrI, 23.5–95.1). High correlation-adjusted specificities were found for both tests (95.0%; [CrI, 91.1–99.6] for the iELISA and 97.7%; [CrI, 95.3–99.4] for the RBT, respectively). The true prevalence of brucellosis was estimated from the serum samples to be 4.6% (95%; [CrI, 0.6–9.5]). The level of agreement between the two tests was evaluated using indices of agreement (n = 995). Good agreement was found for negative results (96.6%; confidence interval [CI], 95.7–97.4), a finding supported by an estimated significant correlation of 0.37 (95%; CI, 0.01–0.73) within the sera testing negative. Agreement was lower for sera testing positive (52.2% CI: 41.9–62.5). The findings highlight the importance of using these two tests in combination as part of any brucellosis control programme.     

Prevalence and risk factors associated with bovine brucellosis in herds under extensive production system in southwestern Nigeria

Using a cross-sectional survey, we determined the prevalence and risk factors associated with bovine brucellosis in herds under extensive production system in southwestern Nigeria. Antibodies to Brucella species in serum samples were tested using the Rose Bengal test (RBT) and competitive enzyme-linked immunosorbent assay (cELISA); for milk, the milk ring test (MRT) and indirect-ELISA (i-ELISA) were used. Questionnaire was administered to cattle herdsmen to determine factors predisposing the animals to bovine brucellosis. Data were analyzed using STATA 12. From 513 serum and 635 milk samples tested among 120 herds, overall animal-level prevalence of 10.1% (95% CI 7.5–12.7%) and 20.2% (95% CI 17.1–23.3%) were recorded by RBT and MRT, respectively; while 9.4% (95% CI 6.9–11.9%) and 17.8% (95% CI 14.8–20.8%) were obtained using cELISA and i-ELISA, respectively. In all, from the 120 herds tested, 29.2% and 43.3% were positive by RBT and MRT, respectively. Multivariable logistic regression revealed that herd location (OR?=?8.12, 95% CI 1.68–38.90) and improper disposal of placenta/fetus (OR?=?17.33, 95% CI 4.81–62.33) were predictors for a seropositive herd using RBT; while herd location (OR?=?5.13, 95% CI 1.27–20.28), large herd size (OR?=?2.62, 95% CI 1.15–5.85), and occurrence of abortion for a year or more (OR?=?4.62, 95% CI 1.53–13.71) were predictors of seropositivity to antibodies to Brucella spp. using MRT. We found high prevalence of brucellosis in cattle herds under extensive management system in southwestern Nigeria. Urgent and coordinated control strategies are required to mitigate this problem.

     

布鲁菌病血清学检测方法的比较

[目的]比较4种布鲁菌病的血清学检测方法。[方法]分别用虎红平板凝集试验（RBT）、间接酶联免疫吸附试验（i-ELISA）、竞争酶联免疫吸附试验（c-ELISA）、试管凝集试验（SAT）4种方法检测876份奶牛血清。[结果]RBT、i-ELISA、SAT、c-ELISA检出的阳性血清分别为543份、501份、460份、500份,阳性率分别是61.99%、57.19%、52.51%、57.08%。