Enzyme-linked immunosorbent assay specific for (1-->6) branched, (1-->3)-beta-D-glucan detection in environmental samples.
(1-->3)-beta-D-Glucans have been recognized as a potential causative agent responsible for bioaerosol-induced respiratory symptoms observed in both indoor and occupational environments. A specific enzyme immunoassay was developed to quantify (1-->6) branched, (1-->3)-beta-D-glucans in environmental samples. The assay was based on the use of a high-affinity receptor (galactosyl ceramide) specific for (1-->3)-beta-D-glucans as a capture reagent and a monoclonal antibody specific for fungal cell wall beta-D-glucans as a detector reagent. The assay was highly specific for (1-->6) branched, (1-->3)-beta-D-glucans (such as that from Saccharomyces cerevisiae) and did not show any response at 200 ng/ml to curdlan, laminarin, pustulan, dextran, mannan, carboxymethyl cellulose, and endotoxins. The detection level was 0.8 ng/ml for baker's yeast glucan and Betafectin. A coefficient of variation of 7.8% was obtained for (1-->3)-beta-D-glucans in house dust samples. Metal working fluids spiked with (1-->3)-beta-D-glucans inhibited the glucan assay. Because the assay is specific for (1-->6) branched, (1-->3)-beta-D-glucans and is sensitive and reproducible, it will be useful for the investigation of health effects from exposure to this class of biologically active molecules.
Milton
D K
DK
Department of Environmental Health, Harvard School of Public Health, Boston, Massachusetts 02115, USA. dmilton@hohp.harvard.edu
Alwis
K U
KU
Fisette
L
L
Muilenberg
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002236:Carbohydrate Conformation; D004285:Dogs; D004391:Dust / Q000032:analysis*; D004731:Endotoxins / Q000032:analysis*; D004783:Environmental Microbiology; D004797:Enzyme-Linked Immunosorbent Assay; D005658:Fungi / Q000737:chemistry*; D005936:Glucans / Q000032:analysis*; D006798:Housing; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D047071:beta-Glucans
D000911:Antibodies, Monoclonal; D004391:Dust; D004731:Endotoxins; D005936:Glucans; D047071:beta-Glucans; C033363:beta-1,3-glucan