A review of enzyme immunoassay for detection of antibody to Brucella abortus in cattle

Nielsen, Klaus; Wright, Peter F.; Kelly, William A.; Cherwonogrodzky, J.H.

doi:10.1016/0165-2427(88)90160-2

Cited by 27 publications

(21 citation statements)

References 65 publications

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“…Serological tests used in the diagnosis of animal brucellosis can be classified, depending on the antigens used, as conventional tests (i.e., those using suspensions of whole cells as antigens) and tests using antigenic extracts. Conventional tests, RBT and CFT in particular, are widely used for cattle and sheep brucellosis (6), but gel precipitation with selected B. abortus and B. melitensis polysaccharides (3,9,10,11,13,14,21,22,27,35) and several ELISAs with S-LPS-rich extracts (3,21,33) have also been proposed. All the above tests have in common that they detect mostly antibodies to antigenic determinants present in the 0 chain of the S-LPS.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of serological tests for diagnosis of Brucella melitensis infection of goats

et al. 1994

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Five serological assays were evaluated for the diagnosis of brucellosis in goats: the rose bengal test (RBT), complement fixation test (CFIT), radial immunodiffusion (RID) with Brucella and Yersinia enterocolitica 0:9 polysaccharides, counterimmunoelectrophoresis (CIEP) with cytosol, and enzyme-linked immunosorbent assay (ELISA) with polyclonal and protein G conjugates and smooth lipopolysaccharide (S-LPS), native hapten polysaccharide (NH), or cytosol antigens. For optimal sensitivity, RBT had to be used with sera-antigen at a 3:1 dilution. In the RID test, Brucella melitensis biotype 1 NH could not be replaced by Brucella abortus biotype 1 or Y. enterocolitica 0:9 polysaccharides. In the ELISA, S-LPS and NH gave similar results and the protein G conjugate increased the specificity. With the sera from 55 B. melitensis culture-positive goats, the sensitivity was 100% for RBT, CFT (titer -4), and ELISA with S-LPS or NH; 94% for RID; and 93% for CIEP. All tests were negative (100% specific) when testing the sera from 127 brucella-free goats. Larger discrepancies among the results of the serological tests were obtained with sera from goats of areas where brucellosis is endemic. When the sera of 20 young goats vaccinated subcutaneously (109 CFU of B. melitensis Rev 1) and bled 6 months later were examined, the specificities were as follows: NH ELISA, 60%; CFT and S-LPS ELISA, 75%;RBT, 80%; CIEP, 90%; and RID, 94%. With the sera from 10 young goats vaccinated conjunctivally (109 CFU of B. melitensis Rev 1) all tests were 100% specific 4 months after vaccination. The proportion of goats giving a positive reaction after vaccination decreased faster in RID than in other tests.

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Section: Discussionmentioning

confidence: 99%

Evaluation of serological tests for diagnosis of Brucella melitensis infection of goats

et al. 1994

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“…To overcome these problems, ELISA procedures were evaluated (reviewed by Nielsen et aL, 1988;Wright et aL, 1990). The indirect format of the ELISA was initially found to have a relatively low specificity compared to the BPAT (Nielsen et aL, 1992).…”

Section: Introductionmentioning

confidence: 99%

The use of divalent cation chelating agents (EDTA/EGTA) to reduce non-specific serum protein interaction in enzyme immunoassay

Nielsen¹,

Kelly²,

Gall³

et al. 1994

Vet Res Commun

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An indirect enzyme immunoassay (ELISA) for detection of bovine antibody to Brucella abortus was modified by the addition of divalent chelating agents to the serum diluent. This addition resulted in an increase in specificity from 96.0% in the regular assay to 99.4% in the modified procedure. Of the 15,715 sera initially tested by the indirect ELISA, 691 that had given positive reactions were selected for retesting in the indirect ELISA with EDTA/EGTA added. The buffered plate antigen test (BPAT) correctly identified 98.6% of the samples as negative. The addition of chelating agents did not alter the sensitivity of the indirect ELISA, which correctly classified 609 sera from animals from which B. abortus had been isolated as positive. The sensitivity of the BPAT was 97.8%.

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“…The BBA tests, the CFT, and the IELISA are affected by antibodies resulting from immunization with Brucella abortus S19, whereas CELISA and the FPA are not significantly affected (2,9,10,15). For goats, the OIE-accepted tests (15) are RBT and the CFT, although they have not been validated using statistical analysis for sheep and goats compared with similar studies of cattle (13); in addition, they have low specificities (probabilities of correctly identifying as negative those animals that are truly negatives [18]) and are affected by antibodies resulting from vaccination of sheep and goats with the Rev.1 strain of Brucella melitensis (3,5).…”

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confidence: 99%

Application of the Fluorescence Polarization Assay for Detection of Caprine Antibodies to Brucella melitensis in Areas of High Prevalence and Widespread Vaccination

Ramírez-Pfeiffer

Nielsen

Smith

et al. 2007

Clin Vaccine Immunol

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The screening Rose Bengal test (RBT), the buffered plate agglutination test (BPAT), and the confirmatory complement fixation test (CFT) are currently approved by the World Organization for Animal Health (OIE) for diagnosis of goat brucellosis. However, RBT (at 3% or 8% cell concentration) is known to be affected by vaccinal antibodies. In the present study, Mexican and Canadian OIE tests were compared with the fluorescence polarization assay (FPA), alone or in combination, using indirect and competitive enzyme-linked immunosorbent assays as classification variables for goat sera obtained from an area of high prevalence and widespread vaccination. The relative sensitivities and specificities were, respectively, 99.7% and 32.5% for RBT3, 92.8% and 68.8% for RBT8, 98.4% and 84.8% for Canadian CFT, 83.7% and 65.5% for Mexican CFT, and 78.1% and 89.3% for FPA. The use of FPA as the confirmatory test in combination with other tests significantly increased the final specificities of the screening tests alone; BPAT, RBT3, and RBT8 plus FPA resulted in final specificities of 90%, 91.2%, and 91.3%, respectively, whereas for the combinations RBT3 plus Mexican CFT, RBT8 plus Mexican CFT, and BPAT plus Canadian CFT, specificities were 65.5%, 63.2%, and 91.7%, respectively. We suggest that FPA may be routinely applied as an adaptable screening test for diagnosis of goat brucellosis and as a confirmatory test for screening test series. Some advantages of FPA are that its cutoff can be adjusted to improve its sensitivity or specificity, it is a low-cost and easy-to-perform test of choice when specificity is relevant or when an alternative confirmatory test is not available, and it is not affected by vaccination, thus reducing the number of misdiagnosed and killed goats.The World Organization for Animal Health (OIE) (15)-approved tests for diagnosis of brucellosis in cattle are the buffered Brucella antigen (BBA) tests (Rose Bengal test [RBT] and buffered plate agglutination test [BPAT]), the complement fixation test (CFT), the indirect (IELISA) and competitive (CELISA) enzyme-linked immunosorbent assays (ELISAs), and the fluorescence polarization assay (FPA). The BBA tests, the CFT, and the IELISA are affected by antibodies resulting from immunization with Brucella abortus S19, whereas CELISA and the FPA are not significantly affected (2, 9, 10, 15). For goats, the OIE-accepted tests (15) are RBT and the CFT, although they have not been validated using statistical analysis for sheep and goats compared with similar studies of cattle (13); in addition, they have low specificities (probabilities of correctly identifying as negative those animals that are truly negatives [18]) and are affected by antibodies resulting from vaccination of sheep and goats with the Rev.1 strain of Brucella melitensis (3, 5). However, it is considered that the high sensitivity (the probability of correctly identifying as positive those animals that are truly positives [18]) of RBT fulfills requirements for surveillance of free areas at the flock level...

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A review of enzyme immunoassay for detection of antibody to Brucella abortus in cattle

Cited by 27 publications

References 65 publications

Evaluation of serological tests for diagnosis of Brucella melitensis infection of goats

Evaluation of serological tests for diagnosis of Brucella melitensis infection of goats

The use of divalent cation chelating agents (EDTA/EGTA) to reduce non-specific serum protein interaction in enzyme immunoassay

Application of the Fluorescence Polarization Assay for Detection of Caprine Antibodies to Brucella melitensis in Areas of High Prevalence and Widespread Vaccination

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