Specific enzyme-linked immunosorbent assay for detection of bovine antibody to Brucella abortus (original) (raw)
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A review of enzyme immunoassay for detection of antibody to Brucella abortus in cattle
Veterinary Immunology and Immunopathology, 1988
Enzyme immunoassay has gained wide acceptance for serological diagnosis of bovine brucellosis because of its ability to detect antibody of all isotypes unlike the conventional tests. The indirect enzyme immunoassay, however, presents several parameters that require careful analysis. These parameters include the choice of antigen and antiglobulin-enzyme conjugate reagents for use in the assay, dealing with the large amount of data the semi-automatic or automatic assay can generate and the inter-and intralaboratory standardization and quality control. This review considers the various methods described in the literature and, briefly, how some of the problems have been overcome or how they might be dealt with.
Journal of Veterinary Diagnostic Investigation, 2011
To detect bovine antibody directed to smooth lipopolysaccharide (LPS), cell lysate (LYS), Opolysaccharide (OPS), and LPS-deprived chromatographic fractions (ChF) of Brucella abortus, 2 bi-antigenic diagnostic models based on the enzymatic rapid immunofiltration assay (ERIFA), ERIFA LPS/LYS and ERIFA OPS/ChF , were developed. Their diagnostic performance was compared with complement fixation test (CFT), Rose Bengal test (RBT), indirect in-house and commercial enzyme-linked immunosorbent assays (iELISA and com-ELISA, respectively), based on the smooth LPS antigen, by using a total of 420 cattle sera collected from aborted-unvaccinated, aborted-unvaccinated and culture-positive, healthy-unvaccinated, and healthy-vaccinated cattle. The results demonstrated excellent agreement and no statistical difference between iELISAs and LPS-, LYS-, OPS-based ERIFA models. However, diagnostic performance of CFT, RBT, and ChF-based ERIFA was less significant than that of LPS-, LYS-, and OPS-based ERIFA models, and iELISAs. The results demonstrated a successful adaptation of the multi-antigenic ERIFA model to anti-B. abortus antibody in bovine sera and suggest that the ERIFA model can be considered as an ''individual rapid ELISA'' due to its similarity with ELISA, individual applicability, and rapidity in determining reactor animals within 5 minutes. In conclusion, the potential of multi-antigenic applications can make the rapid ERIFA model not only an alternative screening method but also a confirmatory test for bovine brucellosis diagnosis.
Journal of clinical microbiology, 1984
A quantitative fluorometric immunoassay (FIAX) was adapted for the detection of serum antibodies to Brucella abortus in cattle. Results are expressed in nanograms of immunoglobulin binding the antigen carrier. The FIAX was compared with the standard tube agglutination, Rivanol precipitation, and complement fixation tests, using 285 serum samples from vaccinated, challenged, or control cattle. Linear regression analysis indicated a significant correlation among all four serological tests; the FIAX test correlated best with the Rivanol test. Ninety sera were from vaccinated and nonvaccinated cattle that were challenged with virulent B. abortus 2308. The sensitivity and specificity of each serological test were determined based on culture results from these cattle. The FIAX was the most sensitive of the four serological tests, detecting 79.2% of the culture-positive animals. The FIAX was the least specific, with 15.4% of the culture-negative animals being classified as positive. Eighty...
Journal of Veterinary Diagnostic Investigation, 2011
To detect bovine antibody directed to smooth lipopolysaccharide (LPS), cell lysate (LYS), Opolysaccharide (OPS), and LPS-deprived chromatographic fractions (ChF) of Brucella abortus, 2 bi-antigenic diagnostic models based on the enzymatic rapid immunofiltration assay (ERIFA), ERIFA LPS/LYS and ERIFA OPS/ChF , were developed. Their diagnostic performance was compared with complement fixation test (CFT), Rose Bengal test (RBT), indirect in-house and commercial enzyme-linked immunosorbent assays (iELISA and com-ELISA, respectively), based on the smooth LPS antigen, by using a total of 420 cattle sera collected from aborted-unvaccinated, aborted-unvaccinated and culture-positive, healthy-unvaccinated, and healthy-vaccinated cattle. The results demonstrated excellent agreement and no statistical difference between iELISAs and LPS-, LYS-, OPS-based ERIFA models. However, diagnostic performance of CFT, RBT, and ChF-based ERIFA was less significant than that of LPS-, LYS-, and OPS-based ERIFA models, and iELISAs. The results demonstrated a successful adaptation of the multi-antigenic ERIFA model to anti-B. abortus antibody in bovine sera and suggest that the ERIFA model can be considered as an ''individual rapid ELISA'' due to its similarity with ELISA, individual applicability, and rapidity in determining reactor animals within 5 minutes. In conclusion, the potential of multi-antigenic applications can make the rapid ERIFA model not only an alternative screening method but also a confirmatory test for bovine brucellosis diagnosis.
PubMed, 1989
Purified O chain of Brucella abortus was passively attached to polystyrene to differentiate antibody responses of cattle vaccinated with B abortus strain 19 from those of naturally infected cattle. In the indirect assay, using O polysaccharide as antigen, a single serum dilution was used and mouse monoclonal antibody to bovine L chain conjugated with horseradish peroxidase was the detection reagent. Measurable antibody was not found in sera of vaccinated cattle, except for 3 sera from cattle that were persistently infected with strain 19. Sera from 25 cattle infected with pathogenic strains contained antibody on the basis of results of indirect enzyme immunoassay, using smooth lipopolysaccharide or O chain as antigens, or results of competitive enzyme immunoassay, using the O-chain antigen. Results in sera from calves with experimentally induced Yersinia enterocolitica serotype 0:9 infection or inoculated with a low dose of B abortus strain 2308 were comparable with those in sera of cattle that were vaccinated with strain 19. The data correlated with those from competitive enzyme immunoassay, using one serum dilution and horseradish peroxidase-conjugated mouse monoclonal antibody to smooth lipopolysaccharide. On the basis of results of the indirect enzyme immunoassay, all sera (except those samples obtained before inoculation) contained antibody to smooth lipopolysaccharide.
Veterinary World, 2021
Background and Aim: The present serodiagnosis of brucellosis in livestock is based on the whole cell or smooth lipopolysaccharide of the Brucella organism in which specificity is hampered by the cross-reactivity, especially with the antibodies against Yersinia enterocolitica O:9 organism. The problem can be addressed by screening for better immunodominant antigens. Hence, the present study was undertaken to screen protein antigens of Brucella abortus for their diagnostic potential in cattle brucellosis. Materials and Methods: Protein antigens of B. abortus (n=10) non-reactive to antibodies against Y. enterocolitica O:9 were selected, expressed in Escherichia coli, assessed the reactivity of expressed recombinant proteins by Western blot, standardized indirect-enzyme-linked immunosorbent assay (ELISA) for detecting Brucella antibodies in cattle serum, and comparative evaluation was done. Results: All the selected protein antigens were expressed and in the Western blot with Brucella antibodies positive cattle serum, six recombinant (Brucella protein 26 [BP26], Cu-Zn Superoxide dismutase [SodC], B. abortus I-1885, Serine protease, Bacterioferritin, and Brucella Lumazine Synthase [BLS]) proteins showed reaction whereas none of the proteins showed reactivity with Brucella negative cattle serum. ELISA has been done using known Brucella positive and negative cattle sera samples (n=113 each) in which the performance of recombinant proteins in diagnosing brucellosis was in the order of BP26 > BLS > SodC followed by rest of the proteins. BP26 based ELISA was found to be better with area under the curve as 0.953, and diagnostic sensitivity, diagnostic specificity, and Youden's index of 90.27%, 95.58%, and 0.8584, respectively, with the excellent agreement (k=0.85). Conclusion: BP26 could be a potential diagnostic antigen among the immunodominant proteins of B. abortus in ruling out Y. enterocolitica O:9 infection while diagnosing brucellosis in cattle herds.
Journal of Veterinary Medicine, Series B, 1989
SummaryA 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 ...
Brazilian Journal of Veterinary Research and Animal Science, 1999
Three soluble antigens were compared by radial immunodiffusion (RID) and agar gel immunodiffusion (AGID) tests: a native haptene (NH) from Brucella melitensis 16M, and a polysaccharide (PS) from B. abortus 1119-3, both obtained by non-hydrolytic methods, and the (O-Chain) polysaccharide extracted also from B. abortus 1119-3 but using an hydrolytic method. Three groups ofbovine scra were tested: a) Naturally infected (n = 76); b) Non-infected (n = 130) and c) S-19 vaccinated (n = 61); the sensitivity (Se), the specificity (Sp) and the ability to differentiate vaccinated (ADV) were determined in each group a, b and c respectively. The highest Se in lhe RID test (84.3%) was achieved by NH; while the three antigens gave 100% Sp. The O-Chain showed 100% ADV in this test. In the AGID test PS antigen showed the best Se (86.6%), and ali antigens showed 100% of Sp and ADV Finally, for its production qualities and efficiency the antigens PS and NH represen~a promising alternative for complementary diagnosis of brucellosis.
2015
Immunochrom atographic test, ELISA. Rapid diagnostic tests are needed to facilitate diagnosis and control of brucellosis. Sero-epidemiology of brucellosis is currently done by employing the Rose Bengal test (RBT). Also the world organization for animal health (OIE) has approved an indirect ELISA for testing serum and milk. The Immuno-Chromatographic brucellosis test (ICT) is a rapid, card-based immunochromatographic test for detection of antibodies directed against B. abortus antigens. To the best of our knowledge, ICT has not been used for the diagnosis of brucellosis in cattle yet. The objective of the study was to evaluate the performance of the ICT brucellosis test for the diagnosis of B. abortus in cattle sera versus RBT and ELISA as a gold standard, Also evaluation of ICT brucellosis test efficacy for detection of antibodies against B. Abortus in milk samples. 94, 90.36 and 84.3 % of sera samples were positive by RBT, ICT and ELISA respectively. The ICT had 94.44 % sensitivity...