Journal of immunological methods · 2025
Development of an ELISA strategy for the serological diagnosis of farmer's lung disease.
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Rouzet Adeline, Devillers Eliane, Barrera Coralie, Scherer Emeline, Millon Laurence, Bellanger Anne-Pauline
Аннотация
CONTEXT: Farmer's lung disease (FLD) is a form of hypersensitivity pneumonitis due to regular exposure to microbial antigens present in moldy hay. Serological diagnosis is usually based on immunoprecipitation techniques using 1 to 12 in-house antigens. To improve standardization, recombinant antigens (r-Ags) have been produced from three etiological agents involved in FLD.
AIM: The aim of this study was to develop a strategy using an ELISA test for the serological diagnosis of FLD.
METHODS: The ELISA strategy was developed on an initial group of patients and then validated using sera from 29 patients with FLD and 54 exposed but non-FLD patients. Using sera from the second group of patients, the ELISA was simplified by using purified antigens from Saccharopolyspora rectivirgula, Lichtheimia corymbifera, and Eurotium amstelodami, each mixed with a corresponding r-Ag. The performance of the ELISA was evaluated according to the antigens used and also associated with an immunoprecipitation technique.
RESULTS: The ELISA test using three purified antigens (PAgs) mixed with 3 r-Ags showed a sensitivity of 97 % and a specificity of 83 % (AUC: 0.84). The use of ELISA as a screening technique and double diffusion (using 6 somatic antigens) as a confirmatory technique in cases of moderate or positive ELISAs showed a sensitivity of 93 % and a specificity of 84 % (AUC: 0.86).
CONCLUSION: The use of a mixture of purified protein antigens and highly standardized recombinant antigens from three etiological agents involved in FLD offers an effective and innovative strategy for the serological diagnosis of FLD cases. This strategy has been successfully applied in the routine workflow of our laboratory.
AIM: The aim of this study was to develop a strategy using an ELISA test for the serological diagnosis of FLD.
METHODS: The ELISA strategy was developed on an initial group of patients and then validated using sera from 29 patients with FLD and 54 exposed but non-FLD patients. Using sera from the second group of patients, the ELISA was simplified by using purified antigens from Saccharopolyspora rectivirgula, Lichtheimia corymbifera, and Eurotium amstelodami, each mixed with a corresponding r-Ag. The performance of the ELISA was evaluated according to the antigens used and also associated with an immunoprecipitation technique.
RESULTS: The ELISA test using three purified antigens (PAgs) mixed with 3 r-Ags showed a sensitivity of 97 % and a specificity of 83 % (AUC: 0.84). The use of ELISA as a screening technique and double diffusion (using 6 somatic antigens) as a confirmatory technique in cases of moderate or positive ELISAs showed a sensitivity of 93 % and a specificity of 84 % (AUC: 0.86).
CONCLUSION: The use of a mixture of purified protein antigens and highly standardized recombinant antigens from three etiological agents involved in FLD offers an effective and innovative strategy for the serological diagnosis of FLD cases. This strategy has been successfully applied in the routine workflow of our laboratory.