A comparison of three serological tests in the diagnosis of caprine brucellosis.

The serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT) were used in the diagnosis of caprine brucellosis. There was a close correlation between the SAT and RBPT when both tests were negative but the RBPT failed to detect 79.82 per cent of sera in excess of 50 iu. Also, owing to the relatively poor milking potential of the Nigerian goat and the false positive results with the MRT, it is concluded that the SAT offers a better serological diagnostic tool for caprine brucellosis in this locality.     

Evaluation of four serological tests for the diagnosis of caprine melioidosis

A complement fixation (CF) test, 2 indirect haemagglutination (IHA-A; IHA-L) tests which differed in antigen preparation and technique, and a microtitre agglutination (MA) test were compared in the serodiagnosis of melioidosis in goats. One hundred and eighteen experimental serums and 3143 field serums from goats in endemic and non-endemic areas of north Queensland were used in the evaluation. Culture of samples for Pseudomonas pseudomallei from 112 goats provided substantiating evidence of infection. The IHA-A test was the most sensitive, and the CF test the most specific. We advocate the use of the IHA-A as a screening test followed by the CF test for confirmation of active melioidosis. The IHA-A test is the better indicator of past infection.     

Evaluation of five serological tests for the diagnosis of porcine brucellosis in French Polynesia

Porcine brucellosis due to Brucella suis biovar 1 raises important issues for pig breeders in French Polynesia. In this region, the disease is enzootic, spreads silently and engenders economic losses in infected farms as well as sporadic human cases. While serological tests are essential in surveillance and control programmes of animal diseases, to date none of the available tests have been shown to be reliable enough to be used as a gold standard in routine individual diagnosis of porcine brucellosis. Few studies about the estimation of the sensitivity and the specificity of porcine brucellosis screening tests have been published, none of them dealing with French Polynesia. The studied population included 1,595 pigs from French Polynesia. Five tests were evaluated: Rose Bengal test, fluorescence polarisation assay, indirect ELISA, and two competitive ELISAs (C-ELISA). The sensitivity and the specificity of each test were estimated. C-ELISA₂ was the most sensitive test (Se C-ELISA₂?=?0.954 [0.889; 0.992] 95 % credibility interval (CrI)) while both C-ELISA and Rose Bengal test (RBT) were the most specific ones (Sp C-ELISA₁?=?0.856 [0.806; 0.915] 95 % CrI; Sp C-ELISA₂?=?0.849 [0.817; 0.879] 95 % CrI; Sp RBT?=?0.853 [0.812; 0.898] 95 % CrI).     

A comparison of standard serological tests for the diagnosis of bovine brucellosis in Canada.

Six agglutination and two complement fixation tests were compared with respect to specificity, sensitivity and relative sensitivity for the serodiagnosis of bovine brucellosis. Based on 1051 sera from brucellosis free herds, the specificity of the tests was 98.9% for the buffered plate antigen test (BPAT), 99.2% and 99.3% for the standard tube and plate agglutination tests (STAT and SPAT), respectively, and 99.8% for the 2-mercaptoethanol test (2MET). On this small sample, the rose bengal plate test (RBPT), card test (CARD) and the complement fixation test (CFT) correctly classed all sera as negative. On a sample of 167 culture positive cattle, the sensitivities of the tests were CFT: 79.0%, BPAT: 75.4, RBPT: 74.9%, CARD: 74.3%, SPAT: 73.1%, STAT: 68.9%, and 2MET: 59.9%. All tests combined detected only 82% of these infected cattle. Analysis of the relative sensitivity of the six agglutination tests gave the following ranking: BPAT greater than RBPT greater than CARD greater than SPAT greater than STAT. The 2MET ranked between the BPAT and RBPT or between the RBPT and CARD depending on the analysis used. The use of the BPAT as a screening test is recommended provided that a test of high specificity and sensitivity such as the CFT is used to confirm screening test reactions.     

Estimation of sensitivity and specificity of five serological tests for the diagnosis of porcine brucellosis

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).     

Development and validation of a competitive ELISA kit for the serological diagnosis of ovine, caprine and bovine brucellosis

A competitive ELISA (Brucella-Ab c-ELISA) was standardized and validated for the detection of Brucella antibodies in cattle, sheep and goat sera using a monoclonal antibody (MAb 4B5A) produced against Brucella melitensis biotype 2. The specificity and sensitivity of the assay were 100% to a 67.5% cut-off point (B/Bo%). When compared with an indirect ELISA, the Brucella-Ab c-ELISA did not demonstrate cross-reactions when testing positive sera for antibodies to some Enterobacteriaceae. A comparison was made between the Brucella-Ab c-ELISA and the complement fixation and Rose Bengal tests. Results demonstrated that the Brucella-Ab c-ELISA is a valuable tool for the serological diagnosis of bovine and ovine/caprine brucellosis.     

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.     

A comparison of five serological tests for bovine brucellosis.

Five serological assays: the buffered plate antigen test, the standard tube agglutination test, the complement fixation test, the hemolysis-in-gel test and the indirect enzyme immunoassay were diagnostically evaluated. Test data consisted of results from 1208 cattle in brucellosis-free herds, 1578 cattle in reactor herds of unknown infection status and 174 cattle from which Brucella abortus had been cultured. The complement fixation test had the highest specificity in both nonvaccinated and vaccinated cattle. The indirect enzyme immunoassay, if interpreted at a high threshold, also exhibited a high specificity in both groups of cattle. The hemolysis-in-gel test had a very high specificity when used in nonvaccinated cattle but quite a low specificity among vaccinates. With the exception of the complement fixation test, all tests had high sensitivities if interpreted at the minimum threshold. However, the sensitivities of the standard tube agglutination test and indirect enzyme immunoassay, when interpreted at high thresholds were comparable to that of the complement fixation test. A kappa statistic was used to measure the agreement between the various tests. In general the kappa statistics were quite low, suggesting that the various tests may detect different antibody isotypes. There was however, good agreement between the buffered plate antigen test and standard tube agglutination test (the two agglutination tests evaluated) and between the complement fixation test and the indirect enzyme immunoassay when interpreted at a high threshold. With the exception of the buffered plate antigen test, all tests were evaluated as confirmatory tests by estimating their specificity and sensitivity on screening-test positive samples.(ABSTRACT TRUNCATED AT 250 WORDS)     

A comparison of four serological tests for the diagnosis of caseous lymphadenitis in sheep and goats

A double antibody sandwich ELISA (ELISA A) developed for the detection of Corynebacterium pseudotuberculosis infection in sheep and goats was modified to improve its sensitivity. To establish the sensitivity and specificity of this modified ELISA (ELISA B), sera from 183 sheep and 186 goats were tested using ELISAs A and B. Comparison was also made with two further ELISAs (C and D) developed in Australia that, respectively, detect antibodies to cell wall antigens or toxin.ELISA B had the best performance of the four tests. Its specificity was 98+/-1% for goats and 99+/-1% sheep. Its sensitivity was 94+/-3% for goats and 79+/-5% for sheep. ELISA B will now be tested for use in caseous lymphadenitis eradication and control programmes in The Netherlands. It will also be used in experimental studies of CL in Scotland.     

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.     

Investigation of aberrant positive reactions to serological tests for bovine brucellosis

Fifty cattle thought not to be infected with Brucella abortus but giving persistent positive serological reactions, were investigated. It was concluded that only one of these was infected since exhaustive bacteriological examination produced only one isolate of B. abortus (strain 19) and none of the herds of origin was subsequently shown to be infected with brucellosis. Antibody was detected in stifle joint fluid of 15 cattle, which may have been stimulated by the presence of Strain 19 antigen that persisted in collagenous tissue long after the viable organisms had been eliminated.     

Comparison of three serological tests in the diagnosis of Brucella infection in unvaccinated cattle in Eritrea

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.     

Comparison and evaluation of four serological tests for detection of antibodies to Aujeszky's disease virus

Four methods used for the detection of antibodies to Aujeszky's disease virus in porcine sera were compared. The serum neutralisation test, countercurrent immunoelectrophoresis test, micro-immunodiffusion test and the enzyme-linked immunosorbent assay (Elisa) were assessed with particular regard to their use in large scale screening of porcine serum samples. The Elisa test was found to be the most suitable on the grounds of sensitivity, speed and cost.     

Equine lentivirus, comparative studies on four serological tests for the diagnosis of equine infectious anaemia.

Serological diagnosis of equine infectious anemia is of necessity group-reactive, i.e. based on viral core protein p26, because viral envelope components as well as the host's immune response to them undergo rapid antigenic change. Since 1970 the agar gel-immunodiffusion test ("Coggins-test") has been the diagnostic method of choice. Recently, ELISA tests have been introduced for faster and theoretically more sensitive serodiagnosis, while Western blots have been used to clarify doubtful results obtained in Coggins-tests. A commercial competitive ELISA was found to give practically equivalent results to the Coggins-test. The sensitivity of this market product is intentionally kept marginal in order to avoid false-positive "reactor horses". Another commercial ELISA, non-competitive, gave inconsistent results, creating great turmoil among horse owners when falsely positive. Caution is also indicated when interpreting Western blots. Sera of strongly positive horses gave as many as eleven bands, of medium positives fewer bands, and of the weakest reactors solely the p26 band. Single p26 banding was, however, also encountered in 5% healthy horses, in two of them consistently over time, which are accordingly considered non-specific. In order to be interpreted as positive, a Western blot for this equine lentivirus must band with its core protein plus at least one glycoprotein, similar to the recommended criterion for a positive reading of serum samples from AIDS patients.     

Comparison of various tests for the serological diagnosis of Trypanosoma equiperdum infection in the horse

Comparative tests such as FAT, ELISA, RIA, IEO and CF in the diagnosis of dourine in the horse have proved a satisfactory concordance ratio of the ELISA with CF, which seems to be the most reliable test. Discrepancies have been observed as to the other tests which appear less sensitive than CF test.