الفهرس | Only 14 pages are availabe for public view |
Abstract Hyaluronidase and coagulase enzymes were obtained from 11 out of 34 studied strains of Staph. aureus isolated from cutaneous abscesses. Hyaluronidase was prepared and purified 150fold from one of the studied strains (Strain No. 117) using cetyl trimethyl ammonium bromide. The specific activity of the enzyme was increased from 750 U/mg to 10<U+00AC>0,000 U/mg. Coagulase enzyme was prepared from the same strain and purified 2100 fold by acid precipitation, DEAE cellulose and ammonium sulfate precipitation. The specific activity of coagulase enzyme increased from 40 U/mg to 85000 U/mg. Rabbit?s antisera were prepared against crude and purified forms of hyaluronidase and coagulase enzymes. The activity of the purified staphylococcal hyaluronidase was neutralized by the specifically prepared antiserum as determined by the viscosityreducing method. The prepared antiserum against crude hyaluronidase produced single and multiple precipitation lines in agarose with purified and crude enzymes isolated from different S. aureus strains, respectively. Antibodies prepared from the purified hyaluronidase revealed a single precipitation line with crude and purified hyaluronidases by using immunodiffusion and immunoelectrophoresis techniques. The prepared antiserum against partially purified coagulase isolated from Staphylococcus aureus strain No. 117 neutralized partially purified coagulases isolated from all the eleven studied strains using enzyme inhibition method, in addition the prepared antiserum was found to produce two precipitation lines in agarose with the partially purified coagulases isolated from the same S. aureus strain as well as coagulases isolated from the other different studied strains in double immunodiffusion assay. These findings conclude that all hyaluronidases and coagulases in this study could be considered as immunologically homogeneous, and the detection of antibodies against hyaluronidase in patient serum may be regarded as an indication of S. aureus infection in a manner similar to detection of antistreptococcal hyaluronidase antiserum. Also antibodies against aminoglycosides inactivating enzymes (AGIEs) were also studied. |