Quantification of [i]C. globosum [/i]spores in house dust samples

[i]Chaetomium globosum [/i]is one of the most common fungi that grows in damp buildings and occurs in agricultural and forestry workplaces. Using sera from atopic patients, we characterized and purified an extracellular chitosanase (Chg47) from [i]C. globosum[/i] that is antigenic to humans. The study reports the production of monoclonal antibodies to the protein. Three capture ELISAs were developed for Chg47 for detection of spores and spore and mycelial fragments in dust samples using different mono- and polyclonal antibody combinations. One method is based on an enhanced biotinylated polyclonal antibody as the secondary antibody and coating anti-IgM to capture one of two clones of IgM monoclonal antibodies as the capture antibody. The other method makes use of an enhanced rabbit polyclonal antibody as both the primary and capture antibody. The detection limit of the double PAb method for the Chg47 antigen was 7.6 pg/ml. When the anti-IgM+10B3 clone was used, the detection limit was 61 pg/ml and for anti-IgM+5F12, 122 pg/ml. The detection limit of double PAb method is comparable to methods for the allergen and spores of [i]Aspergillus versicolor[/i] in house dust and is more sensitive than other immunoassays for allergens in house including for [i]Stachybotrys chartarum[/i], [i]Aspergillus fumigatus[/i] and [i]Alternaria alternata[/i]. All three methods had limited cross-reactivity to fungi common in house dust representing a diverse array of taxa