Differences Between<i>Brucella</i>Antigens Involved in Indirect Hemagglutination Tests with Normal and Tanned Red Blood Cells

Díaz, Ramón; Jones, Lois M.; Leong, Daniel; Wilson, J. B.

doi:10.1128/jb.94.3.499-505.1967

Cited by 12 publications

(9 citation statements)

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“…Immunoelectrophoresis was performed in Noble agar (Difco Laboratories) by using barbital buffer (pH 8.6) with or without 2% Triton X-100 in both the reservoirs and the gel. Indirect hemagglutination with lipopolysaccharide (LPS) was performed as described elsewhere (8). LPS was obtained by the method of Westphal et al (31) from Y. enterocolitica 0:3 grown in Trypticase soy broth (BBL) at 260C.…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Yersinia enterocolitica antigen common to enterocolitis-associated serotypes

Diaz¹,

E²,

Toyos³

et al. 1985

J Clin Microbiol

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Yersinia enterocolitica synthesized an exocellular antigen common to the serotypes associated with enterocolitis but absent from other serotypes or from other Yersinia species. Both virulent Ca2+-dependent and avirulent Ca2+-independent isogenic pairs derived from the enterocolitis-associated serotypes synthesized the common antigen. Requirements for the synthesis of this common antigen were (i) the presence of metabolizable sugars and (ii) growth on a solid medium at 37°C. The antigen was identified as a 24,000-dalton protein loosely associated with the cell surface but absent from either the cell envelope or the cytoplasmic fraction.

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Section: Methodsmentioning

confidence: 99%

Characterization of a Yersinia enterocolitica antigen common to enterocolitis-associated serotypes

Diaz¹,

E²,

Toyos³

et al. 1985

J Clin Microbiol

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“…analyzed by immunoelectrophoresis; the number of components was dependent upon the antigenic preparation and the immune serum employed (7). No relationship was observed between the components in the water extract and the agglutinogen of B. suis or B. abortus (8,20). In contrast, one band of precipitation developed by ether-water extract of B. abortus and B. melitensis when the gel diffusion technique was used could be related to the smooth agglutinogen (8, 10).…”

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confidence: 97%

Surface Antigens of Smooth Brucellae

et al. 1968

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Surface antigens of smooth brucellae were extracted by ether-water, phenol-water, trichloroacetic acid, and saline and examined by immunoelectrophoresis and gel diffusion with antisera from infected and immunized rabbits. Ether-water extracts of Brucella melitensis contained a lipopolysaccharide protein component, which was specific for the surface of smooth brucellae and was correlated with the M agglutinogen of Wilson and Miles, a polysaccharide protein component devoid of lipid which was not restricted to the surface of smooth brucellae and was not correlated with the smooth agglutinogen (component 1), and several protein components which were associated with internal antigens of rough and smooth brucellae. Immunoelectrophoretic analysis of ether-water extracts of B. abortus revealed only two components, a lipopolysaccharide protein component, which was correlated with the A agglutinogen, and component 1. Component 1 from B. melitensis and B. abortus showed identity in gel diffusion tests, whereas component M from B. melitensis and component A from B. abortus showed partial identity with unabsorbed antisera and no cross-reactions with monospecific sera. Attempts to prepare monospecific sera directly by immunization of rabbits with cell walls or ether-water extracts were unsuccessful. Absorption of antisera with heavy fraction of ether-water extracts did not always result in monospecific sera. It was concluded (as has been described before) that the A and M antigens are present on a single antigenic complex, in different proportions depending upon the species and biotype, and that this component is a lipopolysaccharide protein complex of high molecular weight that diffuses poorly through agar gel. Components 1, A, and M were also demonstrated in trichloroacetic acid and phenol-water extracts. With all extracts, B. melitensis antigen showed greater diffusibility in agar than B. abortus antigens. After mild acid hydrolysis, B. abortus ether-water extract was able to diffuse more readily. At least 24 antigenic components were demonstrated in a water extract from Brucella suis

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“…(i) Acetone-killed organisms in incomplete Freund adjuvant were injected according to the procedure previously described (6). (ii) Soluble antigens obtained from living smooth and rough B. abortus strain 11 and suspended in incomplete Freund adjuvant were employed as described (5). (iii) Cell wall antigens were prepared from living B. ovis, rough B. melitensis, rough B. abortus, the dog organism, and acetone-killed smooth B. melitensis.…”

mentioning

confidence: 99%

“…Immunological methods. These have been described previously (5,6). Each serum used in the Ouchterlony plate was tested against 10 double dilutions of antigen, starting with 50 mg per ml.…”

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confidence: 99%

Antigenic Relationship of the Gram-negative Organism Causing Canine Abortion to Smooth and Rough Brucellae

1968

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The use of whole-cell antigens in agglutination and agglutinin-absorption tests showed that the organism causing abortion in dogs is similar to rough Brucella abortus, B. melitensis, and B. ovis, but different from smooth Brucella cultures. Water-soluble antigens obtained by ultrasonic treatment and examined by immunoelectrophoresis and gel diffusion show extensive cross-reactions within the genus Brucella, but little or no cross-reaction with similar antigens from other gramnegative genera in the family Brucellaceae. The dog organism showed near identity with rough and smooth Brucella cultures on the basis of immuno-gel diffusion tests with water-soluble antigens, but it lacked the lipopolysaccharide-endotoxin associated with the agglutinogen of smooth brucellae. These findings support the proposal of Carmichael and Bruner for the designation of a new species, "Brucella canis." The gram-negative organism causing abortion in dogs (1; L. E. Carmichael and D. W. Bruner, Cornell Vet., in press) resembles Brucella suis biotype 3 on a bacteriological basis and rough Brucella cultures on a serological basis (8). The antigenic relationship between rough B. ovis and smooth B. melitensis, as demonstrated by the use of water-soluble antigens in indirect hemagglutination and precipitation tests, has been described (6). Similar analysis of the canine organism, reported here, shows the near identity of this organism with rough and smooth Brucella cultures. Data are also presented to show that soluble antigens of the Brucella species have little or no cross-reaction with similar antigens of some other gram-negative organisms in the family Brucellaceae. MATERIALS AND METHODS Bacterial cultures. Sources or appropriate refferences are given for the following cultures employed: B. melitensis smooth strain 16M and rough strain B115 (6); B. abortus smooth strain 2308, smooth strain 11, rough strain 11 (2); and rough strain 45/20 (11) were obtained from the Bureau of Animal Industry and maintained in the freeze-dried state since 1947. B. ovis strain 0.64.19 (6); and strain REO 198, which does not require added CO2 for growth, was obtained from the National Animal Disease Laboratory, Ames, Iowa. The "dog organism" strain RM 666 was obtained from L.

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Differences BetweenBrucellaAntigens Involved in Indirect Hemagglutination Tests with Normal and Tanned Red Blood Cells

Cited by 12 publications

References 12 publications

Characterization of a Yersinia enterocolitica antigen common to enterocolitis-associated serotypes

Characterization of a Yersinia enterocolitica antigen common to enterocolitis-associated serotypes

Surface Antigens of Smooth Brucellae

Antigenic Relationship of the Gram-negative Organism Causing Canine Abortion to Smooth and Rough Brucellae

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