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. 2002 Jun;40(6):2041-5.
doi: 10.1128/JCM.40.6.2041-2045.2002.

Detection of antibodies specific to an antigenic cell wall galactomannoprotein for serodiagnosis of Aspergillus fumigatus aspergillosis

Che-Man Chan  1 , Patrick C Y Woo, Andy S P Leung, Susanna K P Lau, Xiao-Yan Che, Liang Cao, Kwok-Yung Yuen
Affiliations

Detection of antibodies specific to an antigenic cell wall galactomannoprotein for serodiagnosis of Aspergillus fumigatus aspergillosis

Che-Man Chan et al. J Clin Microbiol. 2002 Jun.

Abstract

Aspergilloma and invasive aspergillosis are important opportunistic infections caused by Aspergillus species, among which Aspergillus fumigatus is the most common species associated with human disease. We developed an enzyme-linked immunosorbent assay (ELISA)-based antibody assay with Afmp1p, a purified recombinant antigenic cell wall galactomannoprotein of A. fumigatus. Evaluation of the test with guinea pig sera against A. fumigatus and other pathogenic fungi indicated that this assay was specific for A. fumigatus. Clinical evaluation revealed that the assay was 100% sensitive for patients with aspergilloma and 33.3% sensitive for patients with invasive aspergillosis. No false-positive results were found for serum samples from 80 healthy blood donors, 6 patients with typhoid fever, 4 patients with melioidosis, 20 patients with penicilliosis marneffei, 5 patients with candidiasis, and 4 patients with cryptococcosis, indicating a high specificity of the test. Thus, this ELISA-based test for the detection of anti-Afmp1p antibody can be of significant value as a diagnostic for aspergillosis.

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Figures

FIG. 1.
FIG. 1.
Western blot analysis of the purified recombinant Afmp1p protein antigen with serum from an A. fumigatus-inoculated guinea pig (lane 1) and preimmune serum (lane 2).
FIG. 2.
FIG. 2.
ELISA-based antibody test detects high levels of specific antibodies only in A. fumigatus- and Afmp1p-inoculated guinea pigs. Serial dilutions of the guinea pig sera were made, and an ELISA was performed to determine the antibody levels in the animal sera. OD405 values were obtained and plotted. One guinea pig was used for each fungus, and three were used for Afmp1p (with the SD indicated).
FIG. 3.
FIG. 3.
Evaluation of sensitivity and specificity of A. fumigatus antibody test of aspergillosis patients. Serum specimens were obtained from 9 patients with culture-documented aspergilloma and from 15 patients with culture-documented invasive aspergillosis. The control serum specimens for the test came from 80 normal blood donors, 6 patients with typhoid fever, 4 patients with melioidosis, 20 patients with penicilliosis marneffei, 5 patients with candidemia, and 4 patients with cryptococcosis. The test results were plotted as OD405s. The cutoff line for positive diagnosis is drawn at a value that equals the sum of the mean value and 5 times the SD of normal blood donors.
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