[Preparation of Staphylococcus aureus antigens for evaluation of their immunological reactivity with the human sera by the western blot method].
Extracellular antigens as well as cell wall extracts of 4 S. aureus strains isolated from different kinds of infection were analysed by Western-Blott technique. Materials obtained in two systems of bacteria cultivation (with and without aeration) were compared. Four systems of PAGE (native conditions, with 8.0 M urea, with SDS and SDS after previous reduction of the material with 2-mercaptoethanol) were compared in order to get the best differentiation of proteins and antigens. Immunological reactivity of the antigens mixture with two human sera: highly positive (with three S. aureus antigens in ELISA) from patient with staphylococcal sepsis and negative (from blood donor) were analysed. The best results were obtained after reduction of the cell wall extracted material in SDS-PAGE. The different protein patterns depending on the strain and the method of bacteria cultivation were observed. The standardisation of Western-Blott technique was performed, including titration of the sera to get the best differentiation of the antigens. The difference in immunological reactivity of the positive and negative sera with staphylococcal antigens mixture showed rather quantitative than qualitative character.
Tyski
S
S
Zakład Bakteriologii PZH w Warszawie.
Jarecka
K
K
Gut
W
W
Hryniewicz
W
W
D000388:Air; D000942:Antigens, Bacterial / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western / Q000379:methods; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D066298:In Vitro Techniques; D008623:Mercaptoethanol / Q000494:pharmacology; D018805:Sepsis / Q000097:blood / Q000276:immunology / Q000382:microbiology*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013203:Staphylococcal Infections / Q000097:blood / Q000276:immunology / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000187:drug effects / Q000254:growth & development / Q000276:immunology*
D000942:Antigens, Bacterial; D003470:Culture Media; D007106:Immune Sera; D012967:Sodium Dodecyl Sulfate; D008623:Mercaptoethanol
Przygotowanie antygenów Staphylococcus aureus do oceny odpowiedzi immunologicznej w surowicy metoda western-blott.