Simultaneous Detection of Antibodies against Apx Toxins ApxI, ApxII, ApxIII, and ApxIV in Pigs with Known and Unknown Actinobacillus pleuropneumoniae Exposure Using a Multiplexing Liquid Array Platform

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Sun, Dong
Hoang, Hai
Yoon, Kyoung-Jin
Halbur, Patrick
Opriessnig, Tanja
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American Society for Microbiology
Giménez-Lirola, Luis
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Veterinary Diagnostic and Production Animal Medicine
The mission of VDPAM is to educate current and future food animal veterinarians, population medicine scientists and stakeholders by increasing our understanding of issues that impact the health, productivity and well-being of food and fiber producing animals; developing innovative solutions for animal health and food safety; and providing the highest quality, most comprehensive clinical practice and diagnostic services. Our department is made up of highly trained specialists who span a wide range of veterinary disciplines and species interests. We have faculty of all ranks with expertise in diagnostics, medicine, surgery, pathology, microbiology, epidemiology, public health, and production medicine. Most have earned certification from specialty boards. Dozens of additional scientists and laboratory technicians support the research and service components of our department.
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Veterinary Microbiology and Preventive Medicine
Our faculty promote the understanding of causes of infectious disease in animals and the mechanisms by which diseases develop at the organismal, cellular and molecular levels. Veterinary microbiology also includes research on the interaction of pathogenic and symbiotic microbes with their hosts and the host response to infection.
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Surveillance for the presence of Actinobacillus pleuropneumoniae infection in a population plays a central role in controlling the disease. In this study, a 4-plex fluorescent microbead-based immunoassay (FMIA), developed for the simultaneous detection of IgG antibodies to repeat-in-toxin (RTX) toxins (ApxI, ApxII, ApxIII, and ApxIV) of A. pleuropneumoniae, was evaluated using (i) blood serum samples from pigs experimentally infected with each of the 15 known A. pleuropneumoniae serovars or with Actinobacillus suis, (ii) blood serum samples from pigs vaccinated with a bacterin containing A. pleuropneumoniae serovar 1, 3, 5, or 7, and (iii) blood serum samples from pigs with an unknown A. pleuropneumoniae exposure status. The results were compared to those obtained in a previous study where a dual-plate complement fixation test (CFT) and three commercially available enzyme-linked immunosorbent assays (ELISAs) were conducted on the same sample set. On samples from experimentally infected pigs, the 4-plex Apx FMIA detected specific seroconversion to Apx toxins as early as 7 days postinfection in a total of 29 pigs inoculated with 14 of the 15 A. pleuropneumoniae serovars. Seroconversion to ApxII and ApxIII was detected by FMIA in pigs inoculated with A. suis. The vaccinated pigs showed poor humoral responses against ApxI, ApxII, ApxIII, and ApxIV. In the field samples, the humoral response to ApxIV and the A. pleuropneumoniae seroprevalence increased with age. This novel FMIA (with a sensitivity of 82.7% and a specificity of 100% for the anti-ApxIV antibody) was found to be more sensitive and accurate than current tests (sensitivities, 9.5 to 56%; specificity, 100%) and is potentially an improved tool for the surveillance of disease and for monitoring vaccination compliance.
This article is published as Giménez-Lirola, Luis G., Yong-Hou Jiang, Dong Sun, Hai Hoang, Kyoung-Jin Yoon, Patrick G. Halbur, and Tanja Opriessnig. "Simultaneous detection of antibodies against Apx toxins ApxI, ApxII, ApxIII, and ApxIV in pigs with known and unknown Actinobacillus pleuropneumoniae exposure using a multiplexing liquid array platform." Clinical and Vaccine Immunology 21, no. 1 (2014): 85-95. DOI: 10.1128/CVI.00451-13. Copyright 2014 American Society for Microbiology. Posted with permission.