Platelia aspergillus gm eia

The GM-EIA assays required 300 μl of serum specimens. The Platelia Aspergillus GM-EIA (Bio-Rad Laboratories, Hungary) was used for GM screening. GM-EIA cut-off values were determined using the OD_450/620 values. While evaluating single specimens any value above the OD (optical density) _450/620 ≧ 0.5 cut-off value was considered positive as requested for in vitro testing.

Platelia Aspergillus GM EIA (Bio-Rad, France) was used to measure the galactomannan of lavage samples according to the manufacturer procedures (15 (link)). Briefly, 300 μl of serum or BAL fluid was added to 100 μl of treatment solution, boiled for three minutes at 104°C and then centrifuged for 10 minutes in 10000× g. Next, 50 μl of supernatant and 50 μl of conjugate were mixed and incubated in microtiter plates precoated with monoclonal antibody EB-A2 for 90 minutes at 37°C. The plates were washed five times; after which they were incubated with 200 μl of tetramethylbenzadine in the dark for 30 minutes. The reaction was stopped by 100 μl of sulfuric acid and absorbance at 450 and 620 nm read using a plate reader. Positive and negative controls were included in each assay. Results were recorded as an index relative to the optical density (OD) of the cut-off control. The GM of lavage and serum was considered positive when OD index was ≥ 0.5. All positive cases were repeated in the same sample before they were considered positive.