Oral fluid as a material for serological diagnostics of caseous lymphadenitis in goats

Abstract Corynebacterium pseudotuberculosis causes caseous lymphadenitis (CLA) in goats and sheep. This study assessed oral fluid (OF) as a minimally invasive sample for CLA serodiagnosis by detecting antibodies to phospholipase D (PLD), CP40 protein, and protein kinase G (PknG) using Western blot (...

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Main Authors: Kinga Biernacka, Marcin Mickiewicz, Zofia Nowek, Agata Moroz-Fik, Magdalena Rzewuska, Ewelina Kwiecień, Małgorzata Murawska, Tomasz Nalbert, Olga Szaluś-Jordanow, Lucjan Witkowski, Iwona Markowska-Daniel, Emilia Bagnicka, Michał Czopowicz, Jarosław Kaba
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-06957-z
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author Kinga Biernacka
Marcin Mickiewicz
Zofia Nowek
Agata Moroz-Fik
Magdalena Rzewuska
Ewelina Kwiecień
Małgorzata Murawska
Tomasz Nalbert
Olga Szaluś-Jordanow
Lucjan Witkowski
Iwona Markowska-Daniel
Emilia Bagnicka
Michał Czopowicz
Jarosław Kaba
author_facet Kinga Biernacka
Marcin Mickiewicz
Zofia Nowek
Agata Moroz-Fik
Magdalena Rzewuska
Ewelina Kwiecień
Małgorzata Murawska
Tomasz Nalbert
Olga Szaluś-Jordanow
Lucjan Witkowski
Iwona Markowska-Daniel
Emilia Bagnicka
Michał Czopowicz
Jarosław Kaba
author_sort Kinga Biernacka
collection DOAJ
description Abstract Corynebacterium pseudotuberculosis causes caseous lymphadenitis (CLA) in goats and sheep. This study assessed oral fluid (OF) as a minimally invasive sample for CLA serodiagnosis by detecting antibodies to phospholipase D (PLD), CP40 protein, and protein kinase G (PknG) using Western blot (WB). Serum and OF samples from 96 goats from a CLA-affected herd in Poland were tested using a commercial ELISA coated with recombinant PLD (rPLD-ELISA; serum only) and WB (serum and OF). Depending on the cut-off, the rPLD-ELISA had 70–80% diagnostic sensitivity. WB on serum showed very high diagnostic sensitivity (98%) and good agreement with the rPLD-ELISA. WB on OF was highly consistent with WB on serum, however it had slightly lower diagnostic sensitivity (90%). Moreover, WB on OF showed substantial variations in antigen detection, with antibodies to PLD being most effectively detected. Considerably fewer OF samples had anti-CP40 and PknG antibodies, likely due to antibody class variations or detection limitations. Further investigation into immunoglobulin classes in different sample types and optimized secondary antibodies is essential for developing an effective OF-based diagnostic test for CLA.
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spelling doaj-art-941e5ea1d6b44ef682401027fb9a1f8c2025-08-20T03:38:15ZengNature PortfolioScientific Reports2045-23222025-07-0115111010.1038/s41598-025-06957-zOral fluid as a material for serological diagnostics of caseous lymphadenitis in goatsKinga Biernacka0Marcin Mickiewicz1Zofia Nowek2Agata Moroz-Fik3Magdalena Rzewuska4Ewelina Kwiecień5Małgorzata Murawska6Tomasz Nalbert7Olga Szaluś-Jordanow8Lucjan Witkowski9Iwona Markowska-Daniel10Emilia Bagnicka11Michał Czopowicz12Jarosław Kaba13Division of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDivision of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDivision of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDivision of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDepartment of Preclinical Sciences, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDepartment of Preclinical Sciences, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDepartment of Preclinical Sciences, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDivision of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDepartment of Small Animal Diseases with Clinic, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDivision of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDivision of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWInstitute of Genetics and Animal Biotechnology, Polish Academy of SciencesDivision of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWDivision of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGWAbstract Corynebacterium pseudotuberculosis causes caseous lymphadenitis (CLA) in goats and sheep. This study assessed oral fluid (OF) as a minimally invasive sample for CLA serodiagnosis by detecting antibodies to phospholipase D (PLD), CP40 protein, and protein kinase G (PknG) using Western blot (WB). Serum and OF samples from 96 goats from a CLA-affected herd in Poland were tested using a commercial ELISA coated with recombinant PLD (rPLD-ELISA; serum only) and WB (serum and OF). Depending on the cut-off, the rPLD-ELISA had 70–80% diagnostic sensitivity. WB on serum showed very high diagnostic sensitivity (98%) and good agreement with the rPLD-ELISA. WB on OF was highly consistent with WB on serum, however it had slightly lower diagnostic sensitivity (90%). Moreover, WB on OF showed substantial variations in antigen detection, with antibodies to PLD being most effectively detected. Considerably fewer OF samples had anti-CP40 and PknG antibodies, likely due to antibody class variations or detection limitations. Further investigation into immunoglobulin classes in different sample types and optimized secondary antibodies is essential for developing an effective OF-based diagnostic test for CLA.https://doi.org/10.1038/s41598-025-06957-zCorynebacterium pseudotuberculosisCLADiagnosticsGoatsImmunoassays
spellingShingle Kinga Biernacka
Marcin Mickiewicz
Zofia Nowek
Agata Moroz-Fik
Magdalena Rzewuska
Ewelina Kwiecień
Małgorzata Murawska
Tomasz Nalbert
Olga Szaluś-Jordanow
Lucjan Witkowski
Iwona Markowska-Daniel
Emilia Bagnicka
Michał Czopowicz
Jarosław Kaba
Oral fluid as a material for serological diagnostics of caseous lymphadenitis in goats
Scientific Reports
Corynebacterium pseudotuberculosis
CLA
Diagnostics
Goats
Immunoassays
title Oral fluid as a material for serological diagnostics of caseous lymphadenitis in goats
title_full Oral fluid as a material for serological diagnostics of caseous lymphadenitis in goats
title_fullStr Oral fluid as a material for serological diagnostics of caseous lymphadenitis in goats
title_full_unstemmed Oral fluid as a material for serological diagnostics of caseous lymphadenitis in goats
title_short Oral fluid as a material for serological diagnostics of caseous lymphadenitis in goats
title_sort oral fluid as a material for serological diagnostics of caseous lymphadenitis in goats
topic Corynebacterium pseudotuberculosis
CLA
Diagnostics
Goats
Immunoassays
url https://doi.org/10.1038/s41598-025-06957-z
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