Detection of galactomannan antigenemia by enzyme immunoassay in experimental invasive aspergillosis.

摘要: A sensitive enzyme immunoassay (EIA) for galactomannan antigenemia that avoids the use of radioisotopes was devised. Three carbohydrate-rich antigenic fractions were purified from Aspergillus fumigatus 2085: a cold alkali extract (CA) mycelium, an acetone-precipitated pyridine (APSK-66) and methanol precipitate culture filtrate. CA APSK-66 further by gel filtration ion-exchange chromatography, respectively. An acid hydrolysate contained only mannose galactose, as determined gas-liquid chromatography. Rabbit antisera raised against conidia, mycelia, cell walls A. fumigatus. By indirect EIA, best immunoglobulin G response (1/8,000) obtained in rabbits immunized intravenously with walls. Antigenemia detected EIA inhibition heat-dissociated sera four immunosuppressed infected but absent two uninfected controls. The circulating antigen resistant to pronase, adsorbed onto concanavalin A, had molecular size 50 100 kilodaltons.