Preparation of antigens from Trichophyton mentagrophytes using a new semi-solid culture medium and their...

A new semi-solid culture medium was developed by substituting the agar in Sabouraud glucose medium by Lutrol FC 127 (BASF, Ludwigshafen, Germany). This culture medium can be liquefied by cooling it down from the incubation temperature to 5-10 degrees C, thus allowing the full harvest of fungal mycelium without any contamination by the gelling agent for antigen preparation. More than 25 protein bands with molecular weights in the range of 98 to 12 kDa were fractionated by SDS-PAGE in antigen preparations from Trichophyton mentagrophytes. Hyperimmune antisera were raised in rabbits and used for immunological studies. Heat-inactivated mycelium was used for the absorption of antibodies against heat-stable cell wall constituents. This absorption facilitated the detection of specific protein bands during immunoblotting which revealed 17 protein antigen bands reacting with antibodies over a range of molecular weights from 98 to 24 kDa.