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Immunoreactive proteins of Saccharopolyspora rectivirgula for farmer's lung serodiagnosis
Coralie Barrera1, Laurence Millon, Bénédicte Rognon
1UMR6249 Chrono-environnement, University of Franche-Comté, Besançon, France; Parasitology-Mycology Department, University Hospital of Besançon, Besançon, France.
Purpose:
Saccharopolyspora rectivirgula is the principal cause of farmer's lung disease (FLD). Serodiagnosis is based on immunoprecipitation techniques or enzyme immunoassays with homemade crude antigens and is not standardized. We aimed to produce specific recombinant antigens for the development of a standardized ELISA.
Experimental Design:
We recruited 41 patients and 43 healthy exposed controls from five university hospital pneumology departments in France and Switzerland. S. rectivirgula proteins were extracted, separated by 2D electrophoresis, and subjected to Western blotting, with sera from FLD patients or controls. FLD-specific proteins were identified by MS and were produced as recombinant antigens. The diagnostic performance of ELISA tests using the recombinant antigens was assessed with all the sera from FLD patients and controls.
Results:
We identified 25 FLD-specific proteins, some of which play important roles in transport, nutrition, or virulence. We produced 17 of these proteins as recombinant antigens and assessed their suitability for inclusion in the ELISA test. A combination of three of these proteins (SR1FA, SR17, and SR22) proved remarkably effective at discriminating between patients and controls, with a sensitivity of 83% and a specificity of 77%.
Conclusions And Clinical Relevance:
The recombinant antigens produced in this study constitute a major step toward the improvement of diagnostic performance and the standardization of FLD serodiagnosis.
Insights
Developing new recombinant antigens for Saccharopolyspora rectivirgula improves farmer's lung disease (FLD) diagnosis. A combination of three antigens demonstrated high sensitivity and specificity in ELISA tests.
Area of Science:
- Respiratory Medicine
- Immunology
- Microbiology
Background:
- Saccharopolyspora rectivirgula is the primary cause of farmer's lung disease (FLD).
- Current serodiagnostic methods for FLD lack standardization and rely on crude antigens.
- There is a need for improved diagnostic tools for FLD.
Purpose of the Study:
- To develop specific recombinant antigens for Saccharopolyspora rectivirgula.
- To create a standardized ELISA for farmer's lung disease serodiagnosis.
Main Methods:
- Proteins from S. rectivirgula were separated using 2D electrophoresis and identified via Western blotting and mass spectrometry.
- FLD-specific proteins were produced as recombinant antigens.
- ELISA tests utilizing these recombinant antigens were evaluated using sera from FLD patients and healthy controls.
Main Results:
- Twenty-five FLD-specific proteins were identified, with 17 produced as recombinant antigens.
- A combination of three recombinant antigens (SR1FA, SR17, SR22) showed significant diagnostic performance.
- The optimized ELISA achieved 83% sensitivity and 77% specificity in distinguishing FLD patients from controls.
Conclusions:
- The developed recombinant antigens represent a significant advancement in FLD serodiagnosis.
- These antigens facilitate improved diagnostic accuracy and standardization of FLD testing.
- This work paves the way for more reliable detection of farmer's lung disease.
