Development of a miniaturized protein microarray as a new serological IgG screening test for zoonotic agents and production diseases in pigs

PLoS One. 2019 May 22;14(5):e0217290. doi: 10.1371/journal.pone.0217290. eCollection 2019.

Abstract

In order to monitor the occurrence of zoonotic agents in pig herds as well as to improve herd health management, the development of new cost-effective diagnostic methods for pigs is necessary. In this study, a protein microarray-based assay for the simultaneous detection of immunoglobulin G (IgG) antibodies against different zoonotic agents and pathogens causing production diseases in pigs was developed. Therefore, antigens of ten different important swine pathogens (Toxoplasma gondii, Yersinia enterocolitica, Salmonella spp., Trichinella spp., Mycobacterium avium, Hepatitis E virus, Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, the porcine reproductive and respiratory syndrome virus, Influenza A virus) were spotted and covalently immobilized as 'antigen-spots' on microarray chips in order to test pig serum for the occurrence of antibodies. Pig serum was sampled at three German abattoirs and ELISA tests for the different pathogens were conducted with the purpose of creating a panel of reference samples for microarray analysis. To evaluate the accuracy of the antigens on the microarray, receiver operating characteristic (ROC) curve analysis using the ELISA test results as reference was performed for the different antigens. High area under curve values were achieved for the antigens of two zoonotic agents: Toxoplasma gondii (0.91), Yersinia enterocolitica (0.97) and for three production diseases: Actinobacillus pleuropneumoniae (0.77), Mycoplasma hyopneumoniae (0.94) and the porcine reproductive and respiratory syndrome virus (0.87). With the help of the newly developed microarray assay, collecting data on the occurrence of antibodies against zoonotic agents and production diseases in pig herds could be minimized to one measurement, resulting in an efficient screening test.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood
  • Antibodies, Protozoan / blood
  • Antibodies, Viral / blood
  • Immunoglobulin G / blood*
  • Mass Screening / methods
  • Mass Screening / veterinary*
  • Miniaturization
  • Protein Array Analysis / methods
  • Protein Array Analysis / veterinary*
  • Serologic Tests / methods
  • Serologic Tests / veterinary
  • Sus scrofa / immunology
  • Swine
  • Swine Diseases / diagnosis*
  • Swine Diseases / immunology
  • Toxoplasma / immunology
  • Trichinella / immunology
  • Yersinia enterocolitica / immunology
  • Zoonoses / diagnosis*
  • Zoonoses / immunology

Substances

  • Antibodies, Bacterial
  • Antibodies, Protozoan
  • Antibodies, Viral
  • Immunoglobulin G

Grants and funding

This work has received funding from the European Innovation Partnership for Agricultural Productivity and Sustainability (EIP-AGRI, https://ec.europa.eu/eip/agriculture/) to DM. The funder provided support in the form of salaries for authors KL and SM. This publication was also supported by Deutsche Forschungsgemeinschaft and University of Veterinary Medicine Hannover, Foundation within the funding programme Open Access Publishing. Both funders did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. Abbott/Alere Technologies GmbH and Indical Bioscience GmbH provided support in the form of salaries for authors RH and CE, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.