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Detection of Invasive Pulmonary Aspergillosis in Haematological Malignancy Patients by using Lateral-flow Technology
Published on: March 22, 2012
Standardization and characterization of antigens for the diagnosis of aspergillosis
Cheila Denise Ottonelli Stopiglia1, Alicia Arechavala, Mariana Carissimi
1Graduate Program in Medicine, Medical Sciences, Laboratory of Pathogenic Fungi, Department of Microbiology, Institute of Basic Health Sciences, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500/Sl 210, CEP 90050-170, Porto Alegre, RS, Brazil.
Abstract:
The aim of this study was to develop and characterize antigens for the diagnosis of aspergillosis. Nine strains of Aspergillus species Aspergillus fumigatus , Aspergillus flavus , and Aspergillus niger were grown in Sabouraud and Smith broth to produce exoantigens. The antigens were tested by immunodiffusion against sera from patients with aspergillosis and other systemic mycoses. The protein fraction of the antigens was detected by SDS-PAGE; Western blot and representative bands were assessed by mass spectrometry coupled to a nano Acquity UltraPerformance LC and analyzed by the Mascot search engine. Concurrently, all sera were tested with Platelia Aspergillus EIA. The most reactive antigens to sera from patients infected by A. fumigatus were produced by A. fumigatus MG2 Sabouraud and pooled A. fumigatus Sabouraud samples, both with a sensitivity of 93% and specificity of 100% and 97%, respectively. Aspergillus niger and A. flavus antigens were reactive against A. niger and A. flavus sera, each one with a sensitivity and specificity of 100%. Two proteins, probably responsible for antigenic activity, β-glucosidase in A. fumigatus and α-amylase in A. niger were attained. The commercial kit had a specificity of 22%, sensitivity of 100%, positive predictive value of 48%, and negative predictive value of 100%. The antigens produced showed high sensitivity and specificity and can be exploited for diagnostics of aspergilloma.
Insights
New antigens for diagnosing aspergillosis were developed and characterized. These novel Aspergillus antigens demonstrate high sensitivity and specificity, offering a promising tool for aspergilloma diagnostics.
Area of Science:
- Medical Mycology
- Diagnostic Immunology
- Protein Chemistry
Background:
- Aspergillosis diagnosis relies on sensitive and specific methods.
- Current diagnostic tools may have limitations in sensitivity or specificity.
- Development of novel antigens is crucial for improved diagnostic accuracy.
Purpose of the Study:
- To develop and characterize novel exoantigens for the diagnosis of aspergillosis.
- To evaluate the diagnostic performance of these antigens against patient sera.
- To identify specific proteins responsible for antigenic activity.
Main Methods:
- Cultivation of Aspergillus species (A. fumigatus, A. flavus, A. niger) to produce exoantigens.
- Immunodiffusion assays using patient sera to test antigen reactivity.
- SDS-PAGE, Western blot, and mass spectrometry for protein characterization.
- Comparison with a commercial Aspergillus enzyme immunoassay (EIA).
Main Results:
- Aspergillus fumigatus antigens showed 93% sensitivity and 100% specificity.
- Aspergillus niger and A. flavus antigens achieved 100% sensitivity and specificity.
- Identified β-glucosidase (A. fumigatus) and α-amylase (A. niger) as potential key antigens.
- Commercial EIA exhibited lower specificity (22%) compared to developed antigens.
Conclusions:
- Developed Aspergillus antigens exhibit high diagnostic sensitivity and specificity.
- These novel antigens are suitable for the diagnosis of aspergilloma.
- The identified proteins (β-glucosidase, α-amylase) are promising targets for future diagnostic development.
