Toxic shock syndrome: modification and comparison of methods for detecting marker proteins in Staphylococcus aureus

Abstract

Development of a new medium and modification of incubation conditions increased production of toxic shock syndrome marker proteins and enabled detection of small volumes of pyrogenic exotoxin C (PEC) by isoelectric focusing and staphylococcal enterotoxin F (SEF) by a newly developed solid-phase radioimmunoassay. The results were compared with those obtained with previously described methods. The results were identical, and all PEC-positive isolates were SEF positive. In a second study of 262 randomly selected Staphylococcus aureus isolates examined by isoelectric focusing and solid-phase radioimmunoassay but grown in fresh beef heart medium, 47 (17.9%) isolates were PEC and SEF positive; however, 9 (3.4%) were PEC positive and SEF negative, and 3 (1.1%) were SEF positive and PEC negative. When grown in buffered beef heart yeast extract medium, six of the previously PEC-positive and SEF-negative isolates were PEC negative. Autoradiographic analysis of selected isolates demonstrated that PEC- and SEF-positive strains bound SEF antitoxin to the protein at isoelectric point 7.2, suggesting that in staphylococci from patients with toxic shock syndrome, PEC and SEF are the same protein. In screening staphylococci for toxic shock syndrome marker proteins, isoelectric focusing to identify PEC may detect false-positive proteins and may be more susceptible to technical variation than immunological methods to detect SEF.