A particle agglutination assay for rapid identification of heparin binding to coagulase-negative staphylococci.
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 micron 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 coagulase-negative staphylococcal strains tested, 30 showed agglutination reactivity with heparin-coated 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, lambda-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.
Pascu
C
C
Department of Medical Microbiology, University of Lund, Sweden.
Hirmo
S
S
Ljungh
A
A
Wadström
T
T
D000372:Agglutination Tests; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000925:Anticoagulants / Q000494:pharmacology; D002351:Carrageenan / Q000494:pharmacology; D003030:Coagulase; D004396:Coloring Agents / Q000494:pharmacology; D003470:Culture Media; D017985:Dalteparin / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D005070:Evans Blue / Q000494:pharmacology; D005079:Excipients / Q000494:pharmacology; D006493:Heparin / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D008163:Luminescent Measurements; D008863:Microspheres; D010504:Periodic Acid / Q000494:pharmacology; D011134:Polysaccharides / Q000494:pharmacology; D013210:Staphylococcus / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D013498:Suramin / Q000494:pharmacology; D013997:Time Factors; D014343:Trypan Blue / Q000494:pharmacology; D014415:Tunicamycin / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D000925:Anticoagulants; D003030:Coagulase; D004396:Coloring Agents; D003470:Culture Media; D005079:Excipients; D011134:Polysaccharides; D010504:Periodic Acid; D014415:Tunicamycin; D005070:Evans Blue; D013498:Suramin; D002351:Carrageenan; D006493:Heparin; C007789:fucoidan; C009288:metaperiodate; D010450:Endopeptidases; D014343:Trypan Blue; D017985:Dalteparin