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.
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.
Authors: Kerri A Johannson; Hayley Barnes; Anne-Pauline Bellanger; Jean-Charles Dalphin; Evans R Fernández Pérez; Kevin R Flaherty; Yuh-Chin T Huang; Kirk D Jones; Leticia Kawano-Dourado; Kevin Kennedy; Melissa Millerick-May; Yasunari Miyazaki; Julie Morisset; Ferran Morell; Ganesh R Raghu; Coreen Robbins; Coralynn S Sack; Margaret L Salisbury; Moises Selman; Martina Vasakova; Simon L F Walsh; Cecile S Rose Journal: Ann Am Thorac Soc Date: 2020-12
Authors: Yadav S Bajagai; Friedrich Petranyi; Sung J Yu; Edina Lobo; Romeo Batacan; Advait Kayal; Darwin Horyanto; Xipeng Ren; Maria M Whitton; Dragana Stanley Journal: Sci Rep Date: 2022-06-30 Impact factor: 4.996
Authors: Ganesh Raghu; Martine Remy-Jardin; Christopher J Ryerson; Jeffrey L Myers; Michael Kreuter; Martina Vasakova; Elena Bargagli; Jonathan H Chung; Bridget F Collins; Elisabeth Bendstrup; Hassan A Chami; Abigail T Chua; Tamera J Corte; Jean-Charles Dalphin; Sonye K Danoff; Javier Diaz-Mendoza; Abhijit Duggal; Ryoko Egashira; Thomas Ewing; Mridu Gulati; Yoshikazu Inoue; Alex R Jenkins; Kerri A Johannson; Takeshi Johkoh; Maximiliano Tamae-Kakazu; Masanori Kitaichi; Shandra L Knight; Dirk Koschel; David J Lederer; Yolanda Mageto; Lisa A Maier; Carlos Matiz; Ferran Morell; Andrew G Nicholson; Setu Patolia; Carlos A Pereira; Elisabetta A Renzoni; Margaret L Salisbury; Moises Selman; Simon L F Walsh; Wim A Wuyts; Kevin C Wilson Journal: Am J Respir Crit Care Med Date: 2020-08-01 Impact factor: 30.528