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Novel Immunofluorescent Technique for Identification of Ovine Immunoglobulins and Other Potential Opsonins Binding to Live Staphylococcus aureus

CSIRO, Division of Animal Health, Pastoral Research Laboratory, Private Mail Bag, P.O., Armidale, New South Wales 2350, Australia

ABSTRACT

A qualitative in vitro technique has been developed for identification of potential opsonins (immunoglobulins G₁, G₂, A, M; the third component of ovine complement C₃, and fibronectin) of sheep origin, binding to cell walls of viable Staphylococcus aureus. The technique uses monospecific antisera to these ovine proteins. The antisera are conjugated to FITC-protein A such that the protein A-binding sites in the Fc region of the immunoglobulin molecules are occupied with FITC-protein A complexes and are prevented, therefore, from binding to protein A in the staphylococcal cell wall. The technique is highly specific and sensitive, and once conjugated monospecific antisera are prepared, many tests can be done in a short time.

Staphylococcus aureus incubated in samples of milk whey showed variable binding of immunoglobulins G₁, G₂, and M; immunoglobulin A was bound from only one sample of milk whey, and the third component of ovine complement C₃ — and fibronectin binding were not detected. Most samples of blood serum and colostral whey produced binding of immunoglobulins G₁, G₂, A, and M, and the third component of ovine complement C₃ to the Staphylococcus aureus cell surface; fibronectin binding was found in serum but not in colostral whey. Washings from involuted mammary glands invariably produced binding of all immunoglobulins but not of the third component of ovine complement C₃ or fibronectin.