Detection of porcine epidemic diarrhea virus–neutralizing antibody using high-throughput imaging cytometry

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Sarmento, Luciana V.
Poonsuk, Korakrit
Tian, Liying
Mora-Díaz, Juan C.
Main, Rodger G.
Baum, David H.
Zimmerman, Jeffrey J.
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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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Porcine epidemic diarrhea virus (PEDV) is an emerging porcine coronavirus that causes a tremendous economic burden on the swine industry. The assessment of PEDV-neutralizing antibody levels provides a valuable tool to assess and predict herd immunity. We evaluated the performance of a PEDV imaging cytometry–based high-throughput neutralization test (HTNT) and compared the HTNT to a fluorescent focus neutralization (FFN) assay using serum samples from pigs of known PEDV infection status (n = 159). Estimates of diagnostic sensitivity and specificity for HTNT and FFN assays derived from receiver-operator characteristic (ROC) curve analyses showed that both PEDV FFN and HTNT provided excellent diagnostic performance. However, in the laboratory, imaging cytometry provided an objective and semi-automated approach that removed human subjectivity from the testing process and reduced the read-time of a 96-well plate to < 4 min. In addition, imaging cytometry facilitated the rapid collection and long-term storage of test images and data for further evaluation or client consultation. For PEDV and other pathogens, imaging cytometry could provide distinct advantages over classic virus neutralization or FFN assays for the detection and quantitation of neutralizing antibody.
This is a manuscript of an article published as Sarmento, Luciana V., Korakrit Poonsuk, Liying Tian, Juan C. Mora-Díaz, Rodger G. Main, David H. Baum, Jeffrey J. Zimmerman, and Luis G. Giménez-Lirola. "Detection of porcine epidemic diarrhea virus–neutralizing antibody using high-throughput imaging cytometry." Journal of Veterinary Diagnostic Investigation 32, no. 2 (2020): 324-328. doi: Posted with Permission. Copyright The Authors 2020.