抗凝剂对布氏杆菌病检测结果的影响

为了解血浆、血清对布氏杆菌病虎红平板凝集试验检测结果是否有影响,试验取20只成年羊的颈静脉血,分别进行了虎红平板凝集试验和试管凝集试验,结果发现：血清样品布氏杆菌病RBPT、SAT的检测结果均为阴性;血浆样品布氏杆菌病RBPT的检测结果均为阳性,SAT的检测结果均为阴性。     

三种血清学检测方法对羊布鲁氏菌病检测对比分析

为客观评价虎红平板凝集试验(RBPT)、试管凝集试验(SAT)和荧光偏振测定法(FPA)对布鲁氏菌病检测的诊断效率和价值,取961份绵羊血清采用三种方法进行检测,检测结果运用卡方检验进行统计学分析。结果表明,RBPT、SAT和FPA的阳性检出率分别为41.31%、14.57%、27.06%,统计学分析结果为P<0.01,三种试验结果均具有统计学意义。RBPT法和FPA法敏感性较好,SAT法特异性较高。在实际检测工作中,应把几种检验方法结合起来应用,并综合流行病学和临床体征分析进行布鲁氏菌病的确诊。     

庆阳市2009～2013年间布鲁氏菌病流行病学调查

基于分析庆阳市畜间布鲁氏菌病（Brucellosis）流行特征、疫情变化趋势及原因,为制定布鲁氏菌病防治对策提供科学评价依据.按照《动物布鲁氏菌病诊断技术》（GB/T 18646～2002）虎红平板凝集试验（RBPT）和试管凝集试验（SAT）操作方法进行采样和检测.共检测羊、牛、猪、犬血清共计209 864份,检出布病阳性血清1415份,阳性率为0.67％.     

试管凝集试验与微量凝集试验检测奶牛布鲁氏菌病效果比较

本文对2021年保存的146份未免疫布病疫苗的奶牛血清进行虎红平板凝集试验,检测出15份虎红抗体阳性,将这15份样品用试管凝集试验(SAT)与微量凝集试验(MSAT)进行复核,两种方法布病抗体阳性均为10份,阳性符合率一致,微量凝集试验多了2份可疑,隔4周后对2只可疑牛只进行重检,一头奶牛转阳,一头变为阴性,两种方法和试剂比对结果一致。相比而言,微量凝集试验具有操作简捷精准、节约试剂耗材、试验结果容易用肉眼辨认、废弃物处置压力小等优势,建议基层兽医实验室推广应用。     

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。     

四种凝集试验检测牛布鲁氏菌病血清抗体的比较

牛布鲁氏菌病主要由流产布鲁氏菌引起的传染病,严重危害着养牛业的发展和人类健康。检测牛布鲁氏菌病血清抗体的方法主要有：试管凝集试验（SAT）、H流基乙醇试管凝集试验（ME—SAT）、虎红平板凝集试验（RBPT）、致敏红细胞凝集试验（SRAT）,平板凝集试验,抗球蛋白凝集试验、补体结合试验、琼脂扩散试验、酶联免疫吸附试验等［’3。作者选用了SAT、ME－SAT、RBPT、SRAT等四种简单常用的方法来检测牛布鲁氏菌病血清抗体,并比较了四种方法的检出率、符合率及优缺点,现将试验结果报告如下。l材料和方法1．l血清正．亚．l待…     

抗凝剂对动物布鲁菌病虎红平板凝集试验的影响

为探讨含抗凝剂样品对动物布鲁菌病虎红平板凝集试验的影响,采集猪、牛、羊等不同动物的血液各10份,经柠檬酸钠、EDTA、肝素钠等抗凝剂处理后与对应的自然凝固、立即离心获得的样品分别进行虎红平板凝集试验和试管凝集试验,比较两种试验结果的差异。结果显示,经抗凝剂处理后的血浆样品虎红平板凝集试验结果均为阳性,试管凝集试验均为阴性,而自然凝固的血清样品两个试验结果均为阴性,不同处理方式造成的虎红平板凝集试验结果差异现象不受动物种类的影响。抗凝剂对虎红平板凝集试验影响非常显著,对试管凝集试验无影响。因此,临床上用虎红平板凝集试验进行动物布鲁菌病筛查时,不能使用含有抗凝剂的血液样品,否则会出现严重的非特异性凝集现象。     

两种常见牛布鲁氏菌病检测方法的比较与分析

为给临床应用提供依据,将牛布鲁氏菌病检测中常用的试管凝集试验（SAT）、捕获酶联免疫吸附试验（C-ELISA）两种方法进行比较和分析。对某奶牛场220份牛血清和60份已知阴阳性样品进行了检测,对检测结果进行Kappa一致性检验并对比敏感性和特异性。结果显示SAT、C-ELISA两种方法具有高度一致性（K=0.732）,SAT的特异性和敏感性（93.33%,86.67%）均低于C-ELISA（96.67%,100%）,表明C-ELISA更适合牛布病的监测,是进行布病诊断、清除和净化工作的重要工具。     

临床就诊犬布鲁氏菌病血清学调查

1绪论犬布鲁氏菌病是由布鲁氏菌侵入机体引起的传染变态反应性人畜共患传染病,国内外均有犬感染该病的报道。犬布鲁氏菌病的检测主要采用国际标准中规定的虎红平板凝集试验(RBPT)、试管凝集试验(SAT)和补体结合试验(CFT)[1]。这些方法具有敏感性高、特异性强的特点。但由于试管凝集试验和补体结合试验操作繁琐、费时,抗原、血清用量较大,试管、吸管反复刷洗,在标本量大的布鲁氏菌病     

共和县布鲁氏菌病的血清学调查

2008-2013年间,应用虎红平板凝集试验（RBT）和试管凝集试验（SAT）对共和县奶牛累计检测布鲁氏菌病的血清7004份,检出阳性血清15份,阳性率0.21%,结果表明共和地区奶牛布鲁氏病得到有效控制。布鲁氏菌病（简称布病）,是由布氏杆菌感染引起的慢性接触性人畜共患的传染病,其特征是妊娠母畜发生流产,公畜呈现睾丸炎。该病可造成奶牛不同程度流产,生产性能下降,     

Comparison of the dot-immunobinding assay with the serum agglutination test, the rose bengal plate test and the milk ring test for the detection of Brucella antibodies in bovine sera and milk.

In this study, Brucella antibodies in bovine sera and milk were detected using the dot-immunobinding assay (DIA), the serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT). For this purpose, a total of 116 paired blood and milk samples collected at the same time from 56 aborted and from 60 healthy dairy cows was examined. In DIA, a nitrocellulose membrane (NCM) was used as the solid phase. Antigen adsorbed on the NCM was extracted from Brucella abortus S99 by heat treatment. The results obtained by DIA were compared with those of SAT, RBPT and MRT. Of the 116 paired blood and milk samples, 24 were positive and 72 were negative by all tests used. Serum samples of six aborted cows were positive by DIA, SAT and RBPT but the milk samples were negative by DIA and MRT. Serum and milk samples of four aborted cows gave positive reaction only by DIA tests. The remaining six aborted cows were negative only by MRT and two of them were negative by both RBPT and MRT. Four sera of healthy cows were found to be positive only by SAT.     

Direct agglutination test for the detection of antibodies to Sarcocystis neurona in experimentally infected animals

Equine protozoal myeloencephalitis (EPM) is a serious neurological disease of horses in the Americas. The apicomplexan protozoan most commonly associated with EPM is Sarcocystis neurona. A direct agglutination test (SAT) was developed to detect antibodies to S. neurona in experimentally infected animals. Merozoites of the SN6 strain of S. neurona collected from cell culture were used as antigen and 2-mercaptoethanol was added to the antigen suspension to destroy IgM antibodies when mixed with test sera. Mice fed sporocysts of S. speeri or S. falcatula-like sporocysts from opossums did not seroconvert in the SAT. The sensitivity of the SAT was 100% and the specificity was 90% in mice.     

A comparison of counter-immunoelectrophoresis with the rose bengal and the serum tube agglutination tests in the diagnosis of brucellosis in sheep

The counter-immunoelectrophoresis test (CIEPT) was compared with the rose bengal plate test (RBPT) and the serum agglutination test (SAT) in the diagnosis of brucellosis in 241 sheep. The total number of animals positive by one or more of the tests used was 106. Sixty animals were positive by all tests, while 87, 79 and 80 were positive by CIEPT, RBPT and SAT respectively. Based on the assumption that animals with an SAT titre of 40 I.U./ml or above were true positives, the CIEPT had a sensitivity of 82.5% and a specificity of 78.3% compared to 96.5% and 87% respectively, demonstrated by the RBPT. CIEPT might be used in flocks without clinical evidence of ram epididymitis if a test to supplement RBPT were required; but it would only partially replace SAT. Sera positive in RBPT but negative in CIEPT would have to be tested by SAT. With this combination, only about 22% of sera positive in RBPT would need to be further tested by SAT.     

细胞壁抗原用于布氏杆菌病诊断试验的研究

用培养的布氏杆菌菌体，通过超声波裂解、反复离心制备出布氏杆菌细胞壁抗原。将细胞壁抗原作1：128稀释，用作牛种布氏杆菌酶联免疫吸附试验（ELISA)的抗原；将布氏杆菌细胞壁抗原作1：32稀释，用作平板凝集试验抗原；把细胞壁抗原作1：16稀释用作试管凝集试验抗原，分别建立了牛布氏杆菌ELISA试验、平板凝集试验和试管凝集试验。用这3种方法，检测已知200份平板凝集试验阴性血清，5份平板凝集试验阳性血清。结果5份阳性血清在平板凝集试验、试管凝集试验和ELISA试验中均为阳性；200份阴性血清在平板凝集试验和试管凝集试验中均为阴性，在ELISA试验中有1份为阳性?试验证明，细胞壁抗原，既能用于传统的平板凝集试验和试管凝集试验，又能用于ELISA试验。     

Reduction of non-specific reactions to the Brucella abortus serum agglutination test by the addition of EDTA

Serum samples taken from cattle known to be infected with Brucella abortus, and samples routinely collected as part of the brucellosis eradication scheme were tested by the serum agglutination test (SAT), complement fixation test (CFT) and the SAT modified by the addition of ethylene diamine tetra-acetic acid (EDTA). In 64 per cent of the samples giving an SAT titre greater than 100 iu, but a CFT titre 8.3 icftu or less, the agglutination reaction was sufficiently affected by the action of EDTA for the titre to drop to below 100 iu. Only 5 per cent of samples giving an SAT titre greater than 100 iu and a CFT titre of 20 icftu or greater were affected in a similar manner by EDTA, and none of the 29 sera taken from known infected animals showed a drop in titre to below 100 iu, although some with titres greater than 500 iu did show significant EDTA sensitivity.     

Comparison of results from five serologic methods used for detecting Brucella abortus antibody activity in coyote sera

A total of 423 serum samples representing 94 coyotes which were wild trapped in east Texas were used to compare the serologic results from five different methods for detecting antibodies to Brucella abortus. The sera were tested for Brucella spp. antibody activity by the Card (CARD), rivanol precipitation (RIV), standard agglutination tube (SAT), cold complement fixation test (CF), and enzyme linked immunosorbent assay (ELISA) methods. Each serum sample selected for this comparison demonstrated antibody activity by one or more of the five serologic methods. When the serologic results of the five different methods were compared, 143 sera were positive according to the CF test and agreement was 67.1-70.6% with CARD, RIV and SAT. The maximum agreement for CF positive was with CARD (70.6%) and the lowest agreement fro CF negative was also with CARD (56.4%). Agreement among the serologic methods for the SAT positive ranged from 69.1% (CARD) to 72.7% (RIV). Agreement between SAT and ELISA was poor with only 38.1% agreement for SAT positive and 11.3% agreement for SAT negative. Agreement between methods for CARD positive sera was poor, with a low of 43% for both SAT and ELISA, and a high of 55.6% for RIV. Agreement between methods for 149 RIV positive sera was 83.2% for CARD, 67.8% for SAT, 64.4% for CF and only 50.3% for ELISA. Agreement between methods for ELISA positive results ranged from 49.0% for RIV to 62.7% for CARD.(ABSTRACT TRUNCATED AT 250 WORDS)     

The relationship between the microtitration serum agglutination and complement fixation tests in bovine brucellosis serology

The relationship between antibody titres in the microtitration serum agglutination test and the complement fixation test in bovine brucellosis is described. For low and high MSAT values there is good agreement between the 2 tests. This is not the case for MSAT values between 54 and 338 IU/ml. For practical reasons, results falling into this category cannot all be repeated. Repetitions are so structured that less than 4% of the tests need to be repeated. If the level of repetitions should show an increase above 4%, it is assumed that technical or human error has occurred.     

Validation of an indirect enzyme-linked immunosorbent assay for the detection of antibody against Brucella abortus in cattle sera using an automated ELISA workstation

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.     

Comparison of dot enzyme-linked immunosorbent assay (dot-ELISA) with other serological tests for the detection of Brucella antibodies in bovine sera

A dot Enzyme-linked Immunosorbent Assay (dot-ELISA), using whole cell Brucella abortus antigen dotted on the nitrocellulose membrane bound to a plastic strip (dipstick) was employed for the detection of Brucella antibodies in bovine sera. The results were compared with that of serum agglutination (SAT), Rose Bengal plate agglutination (RBPT) and Complement Fixation test (CFT). All the four tests gave negative reaction in 127 sera obtained from a brucellosis free herd. Testing of 549 sera from a chronically infected herd revealed 57 positive and 447 negative animals in all the four assays. Of the remaining 45 sera, 34 were positive in dot-ELISA. Six of these cases were independently detected by dot-ELISA while 28 showed positive reactions in combination with other tests. When serum samples from 158 aborted cases were subjected to dot-ELISA, 79 were found positive. Of these dot-ELISA positive cases, 71 gave positive reaction in SAT, 72 in RBPT and 78 in CFT. B. abortus biotype 3 was isolated from 34 of the 98 aborted fetuses examined.     

高原奶牛布鲁氏菌病的血清学调查

为了解青海省民和县奶牛布鲁氏菌病的分布和流行情况,净化布鲁氏菌病,对来自12个乡镇的10308份奶牛血清采用试管凝集反应试验(SAT)进行了血清学检测,检出阳性血清23份,阳性率为0.223%,表明青海民和地区存在奶牛布鲁氏菌的感染。