An improved, rapid competitive ELISA using a novel conserved 3B epitope for the detection of serum antibodies to foot-and-mouth disease virus

Chungwon J. Chung, Alfonso Clavijo, Mangkey A. Bounpheng, Sabena Uddowla, Abu Sayed, Brooke Dancho, Ian C. Olesen, Juan Pacheco, Barbara J. Kamicker, David A. Brake, Carey L. Bandaranayaka-Mudiyanselage, Stephen S. Lee, Devendra K. Rai, Elizabeth Rieder

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

The highly contagious foot-and-mouth disease virus (FMDV) afflicts cloven-hoofed animals, resulting in significant costs because of loss of trade and recovery from disease. We developed a sensitive, specific, and rapid competitive ELISA (cELISA) to detect serum antibodies to FMDV. The cELISA utilized a monoclonal blocking antibody specific for a highly conserved FMDV nonstructural 3B epitope, a recombinant mutant FMDV 3ABC coating protein, and optimized format variables including serum incubation for 90 min at 20–25°C. Samples from 16 animals experimentally infected with one FMDV serotype (A, O, Asia, or SAT-1) demonstrated early detection capacity beginning 7 d post-inoculation. All samples from 55 vesicular stomatitis virus antibody-positive cattle and 44 samples from cloven-hoofed animals affected by non-FMD vesicular diseases were negative in the cELISA, demonstrating 100% analytical specificity. The diagnostic sensitivity was 100% against sera from 128 cattle infected with isolates of all FMDV serotypes, emphasizing serotype-agnostic results. Diagnostic specificities of U.S. cattle (n = 1135) and swine (n = 207) sera were 99.4% and 100%, respectively. High repeatability and reproducibility were demonstrated with 3.1% coefficient of variation in percent inhibition data and 100% agreement using 2 kit lots and 400 negative control serum samples, with no difference between bench and biosafety cabinet operation. Negative results from vaccinated, uninfected cattle, pig, and sheep sera confirmed the DIVA (differentiate infected from vaccinated animals) capability. This rapid (<3 h), select agent–free assay with high sensitivity and specificity, DIVA capability, and room temperature processing capability will serve as a useful tool in FMDV surveillance, emergency preparedness, response, and outbreak recovery programs.

Original languageEnglish (US)
Pages (from-to)699-707
Number of pages9
JournalJournal of Veterinary Diagnostic Investigation
Volume30
Issue number5
DOIs
StatePublished - Sep 1 2018

Bibliographical note

Funding Information:
We thank the USDA-APHIS-FADDL, Greenport, NY for providing FMDV seropositive samples, and Pat Glas, Karen Moran, Andrew Fabian of FADDL for testing samples. We thank Tammy Beckham and Melissa Berquist (Texas A&M University Institute for Infectious Animal Diseases) for guidance and program management, and Sandy Rogers and Carly Ginter (Texas Veterinary Medical Diagnostic Laboratory) for testing bovine negative serum samples. We also thank Scott Adams and Ethan Adams (VMRD) for project coordination. We appreciate Don King and Satya Parida (The Pirbright Institute) for testing some of the negative serum samples. The work of Justin D. Smith and Jos? Barrera (Leidos for U.S. Department of Homeland Security Science and Technology Directorate [DHS S&T], PIADC) for testing the final formatted test kits and for providing sera from vaccinated and infected animals is appreciated. We thank Thomas Burrage (U.S. DHS S&T, PIADC) for initial characterization of mAb 40C8 and Kizzy Bundy (U.S. DHS S&T, Washington, DC) for administrative support. The U.S. Department of Homeland Security Science and Technology Directorate (DHS S&T) provided funding to the Institute for Infectious Animal Diseases (formerly, The National Center for Foreign Animal and Zoonotic Disease Defense) under agreements 2010-ST-061-AG0002, HSHQDC-11-J-00452, HSHQDC-13-J-00269, and HSHQDC-13-J-00241; to the USDA-APHIS-FADDL at PIADC: HSHQDC-09-X-00369; to the USDA Agricultural Research Service Foreign Animal Disease Research Unit at PIADC: HSHQDC-11-X-00189; to Leidos with a subcontract to BioQuest Associates LLC: HSHQDC-09-J-00023 and HSHQDC-14-F-00035; and an unfunded Cooperative Research and Development Agreement between DHS S&T and VMRD. Any opinions contained herein are those of the authors and do not necessarily reflect those of DHS S&T, IIAD, USDA-APHIS-FADDL, USDA-ARS-FADRU, Leidos, BioQuest Associates, or VMRD.

Funding Information:
The U.S. Department of Homeland Security Science and Technology Directorate (DHS S&T) provided funding to the Institute for Infectious Animal Diseases (formerly, The National Center for Foreign Animal and Zoonotic Disease Defense) under agreements 2010-ST-061-AG0002, HSHQDC-11-J-00452, HSHQDC-13-J-00269, and HSHQDC-13-J-00241; to the USDA-APHIS-FADDL at PIADC: HSHQDC-09-X-00369; to the USDA Agricultural Research Service Foreign Animal Disease Research Unit at PIADC: HSHQDC-11-X-00189; to Leidos with a subcontract to BioQuest Associates LLC: HSHQDC-09-J-00023 and HSHQDC-14-F-00035; and an unfunded Cooperative Research and Development Agreement between DHS S&T and VMRD. Any opinions contained herein are those of the authors and do not necessarily reflect those of DHS S&T, IIAD, USDA-APHIS-FADDL, USDA-ARS-FADRU, Leidos, Bio-Quest Associates, or VMRD.

Publisher Copyright:
© 2018 The Author(s).

Keywords

  • DIVA
  • Differentiate infected from vaccinated animals
  • foot-and-mouth disease virus 3ABC protein

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