A particle agglutination assay for rapid identification of heparin binding to coagulase-negative staphylococci

Abstract

The heparin-binding properties of six different species of coagulase-negative staphylococci were examined by a particle agglutination assay. Heparin (mol. wt 4000-6000), mildly treated with sodium periodate, was covalently coupled to amino-modified latex beads (0.72 p m diameter). The particle agglutination assay was validated by comparing results with the adhesion (percentage binding of adherent cells) of coagulase-negative staphylococcal strains to heparinised microtitration plates. Of 38 different coagulasenegative staphylococcal strains tested, 30 showed agglutination reactivity with heparincoated latex beads. Strains of different coagulase-negative staphylococcal species agglutinated heparin-coated latex beads to various extents (e.g., cells of Staphylococcus haemolyticus strains reacted more strongly than cells of S. epidermidis strains). The agglutination reaction was significantly inhibited by fucoidan, suramin, &carrageenan and other sulphated compounds, but not by non-sulphated carbohydrate polymers such as hyaluronic acid. Agglutination of staphylococcal cells with heparin-coated latex beads was completely blocked by a cell-surface extract. These results suggest that structures responsible for heparin binding are exposed on the cell surface.

Heparin Stimulates Staphylococcus aureus Biofilm Formation

“…Consistent with this hypothesis, several groups have evidence suggesting that S. aureus codes for a heparin binding protein (14,16,37). Pascu and colleagues found that 30 of 38 coagulase-negative staphylococci were able to aggregate to heparin-coated beads and this binding was inhibited by several sulfated molecules (48). Another report showed the presence of heparin binding proteins on the surface of Neisseria gonorrhoeae, Helicobacter pylori, S. epidermidis, S. aureus, Staphylococcus pyogenes, and two species of Yersinia (14).…”

Isolation, Selection and Characteristics of Lactobacillus paracasei subsp. paracasei F19

“…Out of \ 400 Lactobacillus strains, the 40 strains expressing the highest binding of porcine mucin were selected for further testing. Mucin binding was repeated with bovine submaxillary gland mucin and porcine type II mucin (Sigma) in a particle agglutination assay (PAA) (6).…”

“…Binding of human bronectin (Fn) (8), collagen type I and III (Collagen Research, Palo Alto, CA, USA), vitronectin (Vn) (9), brinogen (Imco, Stockholm, Sweden, heparin (Fluka Chemie AG, Heidelberg, Germany) and fetuin (Sigma Chemical Co.) was studied in immobilized form at neutral pH and at 3.5 (6).…”

Isolation and characterisation of a 17-kDa staphylococcal heparin-binding protein with broad specificity

“…S. saprophyticus strain TW 111, isolated from a patient with urinary tract infection and previously shown by agglutination assay [9] not to bind heparin was included as a negative control. CNS strains isolated from healthy carriers were also included.…”

“…wts of 66 and 60 kDa, respectively, were earlier identi®ed as binding heparan sulphate (HS) and heparin [7]. Also, CNS may interact with heparin and HS, as shown for whole staphylococcal cells which bound avidly to heparinised polymer surfaces [8], or to heparin immobilised on latex particles [9]. The ability of both S. aureus and CNS strains to bind the heparin-binding growth factors, basic ®broblast growth factor (bFGF) and platelet-derived growth factor (PDGF) has also been shown [10].…”