Abstract: Objective To establish two double-antigen sandwich ELISA systems to detect anti-Afmp1cr and Afmp2cr antibodies
of Aspergillus fumigatus. Methods Recombinant Afmp1cr and Afmp2cr proteins of A.fumigatus expressed in Pichia pastoris were
obtained. Double-antigen sandwich ELISA systems for detecting specific anti-Afmp1cr and anti-Afmp2cr antibodies were
developed after chessboard titrating to determine the appropriate concentrations of the recombinant proteins and HRP-labeled
proteins. The sensitivity of the assay was evaluated using serum samples of rabbits immunized with Afmp1cr and Afmp2cr.
The specificity of the assay was evaluated by detecting serum samples from healthy donors and patients with other pathogenic
fungal and baterial infections. The performance of the two ELISA kits was furthered evaluated using serum samples from
patients with suspected Aspergillus infection. Results The established ELISA kits were capable of detecting anti-Afmp1cr and
anti-Afmp2cr antibodies in immunized rabbit serum at the maximum dilutions of 800 and 3200, respectively. No
cross-reactivity was observed in detecting serum from patients with other pathogenic fungal or bactetial infections. Both of the
two kits yielded positive results in sera from two established Aspergillus-infected cases and a suspected case. Conclusion Two
antibody-capture ELISA kits were developed for the laboratory diagnosis of A.fumigatus infection and can be potentially useful
in the clinical diagnosis of Aspergillosis infections.