Improved methods for detection and serotyping of coagulase from Staphylococcus aureus.
Improved methods for detection and serotyping of staphylocoagulase were concomitantly devised. We devised an improved method for detection of coagulase activity on agarose film in the same manner as single radial immunodiffusion. The amounts of reagents required for detection of coagulase on agarose film were successfully diminished by adding polyethylene glycol (PEG) to the original formula described by Boothby et al. Using microplates in another improved method for coagulase serotyping, the amount of reaction fluid required was considerably less compared with the conventional tube method. PEG was found to be also effective to increase the efficacy of coagulase serotyping. In the presence or absence of anti-coagulase antisera, culture supernatants of staphylococcal strain grown in brain heart infusion broth were incubated with the reaction fluid containing bovine fibrinogen, rabbit plasma, 6-amino caproic acid, polyethylene glycol 6,000. Coagulase activity was visualized as a turbid mass formed in the wells. Turbid mass formation due to coagulase activity was type-specifically inhibited in the presence of type-specific antisera. Detailed procedures of the methods are precisely described with some preliminary results obtained by the methods.
Hwang
S M
SM
Department of Clinical Pathology, Jisan Junior College, Pusan, Korea.
Seki
K
K
Sakurada
J
J
Ogasawara
M
M
Murai
M
M
Ohmayu
S
S
Kurosaka
K
K
Masuda
S
S
D003030:Coagulase / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D003470:Culture Media / Q000032:analysis; D012703:Serotyping / Q000379:methods; D013211:Staphylococcus aureus / Q000145:classification / Q000201:enzymology* / Q000276:immunology
D003030:Coagulase; D003470:Culture Media