Selection of specific immunoreactive proteins of Aspergillus fumigatus from sera of patients with invasive aspergillosis

Objective To screen the protein of Aspergillus fumigatus with the strongest immunoreactivity from the sera of invasive aspergillosis(IA) patients for immunoproteomic analysis.Methods The secreted proteins and mycelial protein of A.fumigatus prepared by TCA/acetone method were coated on ELISA plates.The antibody levels of A.fumigatus in the sera of diagnosed invasive aspergillosis(IA) patients were detected by ELISA.The secreted protein and mycelial protein of A.fumigatus were verified by immunoblot with the positive sera in ELISA.The strongest immunoreactive protein was screened.Results Among the 11 diagnosed IA patients 6 showed positive reaction against A.fumigatus antibody.The immunoblot results indicated that the immunoreactivity of secretory proteins was higher than that of mycelial proteins.Among 4 kinds of culture media,the proteins secreted in yeast peptone glucose medium(YEPG) showed the strongest reactive intensity and the maximum bands.Conclusions The protein secreted by A.fumigatus growing in YEPG for 14 days showed the strongest immunoreactivity and the most maximum bands,so the protein should be useful for further screening the specific antigens of A.fumigatus by immunoproteomic approach.