[The isolation and evaluation of Aspergillus fumigatus antigens].
Antigens from three strains of Aspergillus fumigatus (354, 356, and JIG) and an antiserum against the mixing of these antigens have been produced, and evaluated immunochemically. The antigens were obtained through a modified Coleman & Kaufman technique (culture filtrate concentrated by acetone). Analysis by the immunodiffusion test (ID) against homologous serum has yielded 100% sensitivity (with the studied sera). Concerning heterologous sera we found reactivity with a serum of a patient of candidiasis and another with histoplasmosis. The same result was obtained with a reference antigen in immunodiffusion, showing similar standards of response. Titration of the antiserum by ID and counterimmunoelectrophoresis showed a title of 1:32, and by complement fixation (micro-technique) a title of 1:128. Using immunoelectrophoresis (IEF), the produced antiserum yielded 8 lines of precipitation (5 in the anodic pole and 3 in the cathodic one). In SDS-PAGE at 12.5% the antigen has presented a rather complex electrophoretic profile (26 proteic subunits with a molecular weight ranging from 18 a > 100 kDa). Immunogenicity of the antigen was observed in all fractions of SDS-PAGE when the immunoblotting against the antiserum was carried out.
Lirio
V de S
Vde S
Laboratório de Micologia Médica, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, Brasil.
de Assis
C M
CM
Cano
M I
MI
Lacaz
C da S
Cda S
D000818:Animals; D000946:Antigens, Fungal / Q000032:analysis / Q000302:isolation & purification*; D001228:Aspergillosis / Q000175:diagnosis; D001232:Aspergillus fumigatus / Q000276:immunology*; D006801:Humans; D007106:Immune Sera / Q000032:analysis / Q000302:isolation & purification; D007114:Immunization; D007120:Immunochemistry; D007158:Immunologic Techniques; D008297:Male; D011817:Rabbits
D000946:Antigens, Fungal; D007106:Immune Sera
Obtenção e avaliação de antígenos de Aspergillus fumigatus.