Radial immunodiffusion test with a Brucella polysaccharide antigen for differentiating infected from vaccinated cattle

Díaz, Ramón; Garatea, P; Jones, Lois M.; Moriyón, Ignacio

doi:10.1128/jcm.10.1.37-41.1979

Cited by 85 publications

(66 citation statements)

References 3 publications

(4 reference statements)

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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%

“…Therefore, the lower sensitivity reported before for RBT and CFT (18,40) is possibly due both to the use of sera from animals of unknown infectious status and, for the RBT, to the use of the standard method. Also, the RID test described for cattle (11,13,14,22) had to be modified in two ways. First, a lower polysaccharide concentration was necessary (5 ,ug/ml versus the 15 to 20 ,ug/ml used for cattle [13,14]), suggesting that the average antibody response of goats to the B. melitensis M and C epitopes is somewhat lower than that of cattle to the A and C epitopes of B. abortus (15).…”

Section: Discussionmentioning

confidence: 99%

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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%

Section: Discussionmentioning

confidence: 99%

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

et al. 1994

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“…The development of an effective subcellular vaccine devoid of lipopolysaccharide (LPS) might resolve the single greatest obstacle to eradication: detection of latently infected animals (43). Additionally, diagnostic tests not reliant on the measure of LPS antibody would be a highly desirable alternative, although progress has been made in distinguishing vaccination from infection titers by decreasing the vaccine 979 dose (3) and by the observation that most infected cattle produce a response to the native hapten of LPS not detectable in vaccinated animals (16,39). Although the proteins under investigation are structural constituents of the cell, and therefore unlikely to function primarily as virulence factors, they may induce immune responses useful in diagnosis and in conferring protective immunity.…”

mentioning

confidence: 99%

“…Trichloroacetic acid (TCA) extractions of whole cells, or of cytoplasmic constituents derived from the 150,000 x g supernatant after pressure cell disruption, were performed by the method of Diaz et al (16) and hydrolysis of TCA extracts by the method of Moreno et al (39).…”

mentioning

confidence: 99%

Outer membrane proteins of Brucella abortus: isolation and characterization

Verstreate¹,

Creasy²,

Caveney³

et al. 1982

Infect Immun

147

124

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Outer membrane proteins were derived from one rough and four smooth strains of Brucella abortus by sequential extraction of physically disrupted cells with N-lauroylsarcosinate and dipolar ionic detergent. Extraction of outer membrane proteins was ineffective, however, without predigestion with lysozyme. Three groups of proteins were present and could be separated in their native state by sequential anion-exchange chromatography and gel filtration. Membrane proteins contained substantial quantities of tightly adherent lipopolysaccharide which could be reduced but not eliminated by extraction of cells with trichloroacetic acid before disruption. Group 2 proteins, apparently trimers in their native state, gave rise to 43,000- and 41,000-molecular-weight bands after complete denaturation in sodium dodecyl sulfate. They were antigenically identical among all the strains, showed close resemblance in amino acid composition to each other and a general similarity to OmpF of Escherichia coli, and are proposed to be the porins of B. abortus. Group 3 proteins occurred as 30,000-molecular-weight bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, although additional bands were frequently observed in this region. In none of the strains did group 3 proteins manifest heat-modifiable characteristics. Proteins of different strains bore a high degree of similarity to each other in amino acid composition, except in methionine, isoleucine, tyrosine, and histidine. Differences occurred consistently in amino acid composition between group 2 and 3 proteins, and some of these correspond to differences between OmpF and OmpA. Group 2 and 3 proteins were antigenically distinct from each other, but the principal group 3 antigens were shared among all the strains. Despite the lack of heat modifiability, perhaps influenced by adherent lipopolysaccharide, group 3 proteins are proposed as counterparts to OmpA. Most of the group 1 proteins, minor components, were physically associated with those of group 3 unless in sodium dodecyl sulfate. Group 1 proteins produced a major band at 94,000 and exhibited heat modifiability. No evidence was found of a low-molecular-weight lipoprotein in the outer membrane of B. abortus, but this is not taken to exclude its occurrence.

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“…Poly B has received considerable interest because B. abortus infected animals produce antibodies which precipitate this carbohydrate while vaccinated animals do not (Diaz et al, 1979;Jones et al, 1980). Its composition is unclear; it is chemically different from native hapten (NH) or acid hapten (AH) (Moreno et al, 1981;Perera et al, 1984); it is purified from rough rather than smooth strains , it is made within the cell of rough strains and it is immunologically distinct as determined by its inability to absorb out all antibodies to NH (Moreno et al, 1981 ).…”

Section: Brucella Antigensmentioning

confidence: 99%

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

Nielsen

Wright

Kelly

et al. 1988

Veterinary Immunology and Immunopathology

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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.

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Radial immunodiffusion test with a Brucella polysaccharide antigen for differentiating infected from vaccinated cattle

Cited by 85 publications

References 3 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

Outer membrane proteins of Brucella abortus: isolation and characterization

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

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