de Repentigny L, Boushira M, Ste-Marie L, Bosisio G
J Clin Microbiol. 1987 May;25(5):863-7. doi: 10.1128/jcm.25.5.863-867.1987.
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) from mycelium, an acetone-precipitated pyridine extract (APSK-66) from mycelium, and a methanol precipitate from culture filtrate. CA and APSK-66 were further purified by gel filtration and ion-exchange chromatography, respectively. An acid hydrolysate of CA contained only mannose and galactose, as determined by gas-liquid chromatography. Rabbit antisera were raised against conidia, mycelia, and cell walls of A. fumigatus. By indirect EIA, the best immunoglobulin G response (1/8,000) was obtained against CA in rabbits immunized intravenously with cell walls. Antigenemia was detected by indirect EIA inhibition in heat-dissociated sera of four immunosuppressed rabbits that were infected intravenously but was absent in two uninfected controls. The circulating antigen was resistant to pronase, was adsorbed onto concanavalin A, and had a molecular size of 50 to 100 kilodaltons.
设计了一种用于检测半乳甘露聚糖抗原血症的灵敏酶免疫测定法（EIA），该方法无需使用放射性同位素。从烟曲霉2085中纯化出三种富含碳水化合物的抗原组分：菌丝体的冷碱提取物（CA）、菌丝体的丙酮沉淀吡啶提取物（APSK-66）以及培养滤液的甲醇沉淀物。CA和APSK-66分别通过凝胶过滤和离子交换色谱进一步纯化。通过气液色谱法测定，CA的酸水解产物仅含有甘露糖和半乳糖。用烟曲霉的分生孢子、菌丝体和细胞壁制备兔抗血清。通过间接EIA，在用细胞壁静脉免疫的兔子中，针对CA获得了最佳的免疫球蛋白G反应（1/8,000）。通过间接EIA抑制法在四只静脉感染的免疫抑制兔子的热解离血清中检测到抗原血症，但在两只未感染的对照中未检测到。循环抗原对链霉蛋白酶有抗性，能被伴刀豆球蛋白A吸附，分子大小为50至100千道尔顿。
