Staphylococcus aureus exopolysaccharide in vivo demonstrated by immunomagnetic separation and electron microscopy

Johne, Berit; Jarp, J.; Haaheim, Lars R.

doi:10.1128/jcm.27.7.1631-1635.1989

Cited by 58 publications

(27 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Gel negatively-stained with Coomassie blue. Lane: 1: Bio-Rad prestained broad range MW standards; 2: Phdp ECPs produced in brain heart infusion broth 2% NaCl; 3: sea bass abdominal washings; 4: Phdp ECPs isolated after 92 h insertion in the abdominal cavity of sea bass; 5: 98 h; 6: 117 h; 7: 140 h; 8: 210 h. capsule by Staphylococcus aureus (Johne, Jarp & Hanson 1989) and Mycoplasma dispar (Almeida & Rosenbusch 1991) and the expression of novel protein antigens by Campylobacter jejuni (Panigrahi, Losonsky, DeTolla & Morris 1992) and Aeromonas salmonicida (Thornton, Garduno, Carlos & Kay 1993). The occurrence and importance of this phenomenon has been reviewed by Brown, Anwar & Costerton (1988) and Smith (1990).…”

Section: Discussionmentioning

confidence: 99%

The effect of in vivo growth on the cellular and extracellular components of the marine bacterial pathogen Photobacterium damsela subsp. piscicida

Bakopoulos¹,

Hanif²,

Πούλος³

et al. 2004

Journal of Fish Diseases

View full text Add to dashboard Cite

Photobacterium damsela subsp. piscicida, the causative agent of fish pasteurellosis, was grown in vivo. Bacterial cells and extracellular products (ECPs) were analysed via electrophoresis and immunoblot analysis, using specific sea bass antisera. Growth in vivo induced the synthesis of unique bacterial cell proteins at > 206, 206, 21.3, 18, 7.6 and < 7.6 kDa. Sea bass serum raised against live bacterial cells of the pathogen and especially a sea bass serum raised against formalin-inactivated bacterial cells grown in a specific novel medium recognized the novel antigens at > 206 (associated with iron sequestration), 21.3, 7.6 and < 7.6 kDa, suggesting that the latter medium conserves the synthesis of natural bacterial cell proteins in vitro. In vivo growth of the pathogen induced the synthesis of more toxic ECPs in comparison with in vitro growth and an inverse correlation between total protein concentration in the ECPs and toxicity per unit of protein was observed. Substrate-polyacrylamide electrophoresis revealed the presence of in vivo synthesized ECPs of the pathogen (proteases) at 175, 132, < 79 and 48.3 kDa. Histological examination of tissues isolated from fish injected with these ECPs revealed inflammatory and necrotic lesions in the spleen, liver, head kidney, intestine and heart as soon as 48 h post-introduction of the ECPs.

show abstract

Section: Discussionmentioning

confidence: 99%

The effect of in vivo growth on the cellular and extracellular components of the marine bacterial pathogen Photobacterium damsela subsp. piscicida

Bakopoulos¹,

Hanif²,

Πούλος³

et al. 2004

Journal of Fish Diseases

View full text Add to dashboard Cite

show abstract

“…Immunomagnetic beads have been successfully used for the isolation of bacterial cells or their products in agricultural (Jones and Van Vuurde 1989) and medical research (Cudjoe et al 1991). Material adsolbed to magnetic beads was analysed by dilution plating methods (Vermunt et al 1992), ELISA (Gunderson et al 1992), electron microscopy (Johne et al 1989) and polymerase chain reaction .…”

Section: Discussionmentioning

confidence: 99%

Verification of ELISA results by immunomagnetic isolation of antigens from extracts and analysis with SDS‐PAGE and Western blotting, demonstrated for Erwinia spp. in potatoes

Wolf¹,

Beckhoven²,

Vries³

et al. 1994

Journal of Applied Bacteriology

View full text Add to dashboard Cite

Isolation of antigens on immunomagnetic beads and subsequent analysis with SDS-PAGE and Western blotting (immunomagnetic isolation-Western blotting (IMI-WB)) was used to verify positive ELISA results for Erwinia chrysanthemi and Erw. carotovora subsp. atroseptica in potato peel extracts. Direct analysis of highly contaminated extracts by Western blotting without previous immuno-isolation resulted in background reactions, whereas immunomagnetic isolation resulted in distinct bands of specific antigens. Target cells as well as antigenic cell products were captured in IMI-WB. Band patterns on IMI-WB of cell-free culture filtrates and cell suspensions were highly similar, but the removal of cells lowered the detection level by 10- to 100-fold. Threshold levels of IMI-WB were generally comparable with those of ELISA. No differences in threshold levels and band patterns were found between a direct format and an indirect format of immuno-isolation. In IMI-WB, blotting patterns differed between Erw. chrysanthemi and Erw. carotovora subsp. atroseptica. The patterns were identical for 15 Erw. chrysanthemi strains, isolated from potato peel extracts in The Netherlands. However, one of 15 strains of Erw, carotovora subsp. atroseptica from potato peel extracts in The Netherlands gave an aberrant pattern. Target bacteria could be easily distinguished from those of cross-reacting strains on the basis of band patterns. Potato peel extracts naturally contaminated with Erw. chrysanthemi gave IMI-WB patterns that were similar to pure cultures of the homologous strains.

show abstract

“…In a pulse-chase experiment, cell-bound CPS was released continuously into the culture fluid at the same rate as cell wall turnover and there was no evidence of direct excretion of CPS. from a wide range of human and animal sources, and have been demonstrated in vivo in cases of bovine mastitis [1]. Unambiguous evidence that the capsule is a significant virulence factor in mice has come from studies of transposon-induced mutants with reduced levels of capsular polysaccharide (CPS) [2].…”

Section: Discussionmentioning

confidence: 99%

Turnover of cell surface-bound capsular polysaccharide in Staphylococcus aureus

Hancock

1991

FEMS Microbiology Letters

View full text Add to dashboard Cite

The capsular polysaccharide (CPS) of Staphylococcus aureus strain Smith was labelled by growth of bacteria in the presence of radioactive N-acetylglucosamine and was separated from labelled cell wall components by affinity chromatography on wheat germ agglutinin following dissolution of the cells by lysostaphin. The products were partially characterised chemically and immunochemically. Similar labelled components were found in the culture fluid during growth. In a pulse-chase experiment, cell-bound CPS was released continuously into the culture fluid at the same rate as cell wall turnover and there was no evidence of direct excretion of CPS.

show abstract

Staphylococcus aureus exopolysaccharide in vivo demonstrated by immunomagnetic separation and electron microscopy

Cited by 58 publications

References 27 publications

The effect of in vivo growth on the cellular and extracellular components of the marine bacterial pathogen Photobacterium damsela subsp. piscicida

The effect of in vivo growth on the cellular and extracellular components of the marine bacterial pathogen Photobacterium damsela subsp. piscicida

Verification of ELISA results by immunomagnetic isolation of antigens from extracts and analysis with SDS‐PAGE and Western blotting, demonstrated for Erwinia spp. in potatoes

Turnover of cell surface-bound capsular polysaccharide in Staphylococcus aureus

Contact Info

Product

Resources

About