1887

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Volume 71, Issue 4

Development of an ELISA to distinguish between foot-and-mouth disease virus infected and vaccinated animals utilising the viral non-structural protein 3ABC Open Access

Abstract

Foot-and-mouth disease (FMD) is a highly contagious and economically devastating viral disease of livestock and is endemic in much of Asia, including Pakistan. Vaccination is used to control disease outbreaks and sensitive diagnostic methods which can differentiate infected animals from vaccinated animals (DIVA) are essential for monitoring the effectiveness of disease control programmes. Tests based on the detection of the non-structural protein (NSP) 3ABC are reliable indicators of virus replication in infected and vaccinated populations.

Diagnosis of FMD is expensive using commercial ELISA kits, yet is essential for controlling this economically-important disease.

The development of a low-cost diagnostic ELISA, using protein made in .

In this study, the viral precursor protein 3ABC (r3ABC) was expressed in , solubilised using detergent and purified using nickel affinity chromatography. The fusion protein contained an attenuating mutation in the protease and a SUMO tag. It was characterised by immunoblotting and immunoprecipitation, which revealed antigenicity against virus-specific polyclonal sera. Using r3ABC, an indirect ELISA was developed and evaluated using field sera from healthy/naïve, vaccinated and infected animals.

The diagnostic sensitivity and specificity of the r3ABC in-house ELISA were 95.3 and 96.3% respectively. The ELISA was validated through comparison with the commercially available ID Screen FMD NSP competition kit. Results indicated good concordance rates on tested samples and high agreement between the two tests.

The ELISA described here can effectively differentiate between infected and vaccinated animals and represents an important low cost tool for sero-surveillance and control of FMD in endemic settings.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): 3ABC , diagnosis , ELISA , FMD , foot-and-mouth disease and uncleavable precursor
Funding
This study was supported by the:
  • Pakistan Atomic Energy Commission (Award Technical Cooperation Project RAS5085)
    • Principle Award Recipient: MudasserHabib
  • Higher Education Commision, Pakistan (Award 1-8/HEC/HRD/2019/8802/PIN: IRSIP 43 BMS 96)
    • Principle Award Recipient: MuhammadAshir Zia
  • Biotechnology and Biological Sciences Research Council (Award BB/T015748/1)
    • Principle Award Recipient: DavidJ. Rowlands
  • Biotechnology and Biological Sciences Research Council (Award BB/T015748/1)
    • Principle Award Recipient: SamuelJ. Dobson
  • Biotechnology and Biological Sciences Research Council (Award BB/T015748/1)
    • Principle Award Recipient: NicolaJ. Stonehouse
© 2022 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2022-04-06
2024-04-24
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References

  1. Gao Y, Sun S-Q, Guo H-C. Biological function of foot-and-mouth disease virus non-structural proteins and non-coding elements. Virol J 2016; 13:1–17 [View Article]
     [Google Scholar]
  2. Belsham G. Translation and replication of FMDV RNA. Foot-and-Mouth Disease Virus 200543–70
     [Google Scholar]
  3. Mason PW, Grubman MJ, Baxt B. Molecular basis of pathogenesis of FMDV. Virus Res 2003; 91:9–32 [View Article] [PubMed]
     [Google Scholar]
  4. Jamal SM, Belsham GJ. Foot-and-mouth disease: past, present and future. Vet Res 2013; 44:1–14 [View Article] [PubMed]
     [Google Scholar]
  5. Farooq U, Ahmed Z, Naeem K, Bertram M, Brito B et al. Characterization of naturally occurring, new and persistent subclinical foot-and-mouth disease virus infection in vaccinated Asian buffalo in Islamabad Capital Territory, Pakistan. Transbound Emerg Dis 2018; 65:1836–1850 [View Article] [PubMed]
     [Google Scholar]
  6. Elnekave E, van Maanen K, Shilo H, Gelman B, Storm N et al. Prevalence and risk factors for foot and mouth disease infection in small ruminants in Israel. Prev Vet Med 2016; 125:82–88 [View Article] [PubMed]
     [Google Scholar]
  7. Robiolo B, Seki C, Fondevilla N, Grigera P, Scodeller E et al. Analysis of the immune response to FMDV structural and non-structural proteins in cattle in Argentina by the combined use of liquid phase and 3ABC-ELISA tests. Vaccine 2006; 24:997–1008 [View Article] [PubMed]
     [Google Scholar]
  8. Barnett PV, Geale DW, Clarke G, Davis J, Kasari TR. A review of OIE country status recovery using vaccinate-to-live versus vaccinate-to-die foot-and-mouth disease response policies I: benefits of higher potency vaccines and associated NSP DIVA test systems in post-outbreak surveillance. Transbound Emerg Dis 2015; 62:367–387 [View Article] [PubMed]
     [Google Scholar]
  9. Rweyemamu M, Roeder P, MacKay D, Sumption K, Brownlie J et al. Planning for the progressive control of foot-and-mouth disease worldwide. Transbound Emerg Dis 2008; 55:73–87 [View Article] [PubMed]
     [Google Scholar]
  10. Clavijo A, Wright P, Kitching P. Developments in diagnostic techniques for differentiating infection from vaccination in foot-and-mouth disease. Vet J 2004; 167:9–22 [View Article] [PubMed]
     [Google Scholar]
  11. Biswal JK, Jena S, Mohapatra JK, Bisht P, Pattnaik B. Detection of antibodies specific for foot-and-mouth disease virus infection using indirect ELISA based on recombinant nonstructural protein 2B. Arch Virol 2014; 159:1641–1650 [View Article] [PubMed]
     [Google Scholar]
  12. Mahajan S, Mohapatra JK, Pandey LK, Sharma GK, Pattnaik B. Truncated recombinant non-structural protein 2C-based indirect ELISA for FMD sero-surveillance. J Virol Methods 2013; 193:405–414 [View Article] [PubMed]
     [Google Scholar]
  13. Biswal JK, Ranjan R, Pattnaik B. Diagnostic application of recombinant non-structural protein 3A to detect antibodies induced by foot-and-mouth disease virus infection. Biologicals 2016; 44:157–162 [View Article] [PubMed]
     [Google Scholar]
  14. Fu Y, Li P, Cao Y, Wang N, Sun P et al. Development of a blocking ELISA using a monoclonal antibody to a dominant epitope in non-structural protein 3A of foot-and-mouth disease virus, as a matching test for a negative-marker vaccine. PLoS One 2017; 12:e0170560 [View Article] [PubMed]
     [Google Scholar]
  15. Yang M, Parida S, Salo T, Hole K, Velazquez-Salinas L et al. Development of a competitive enzyme-linked immunosorbent assay for detection of antibodies against the 3B protein of foot-and-mouth disease virus. Clin Vaccine Immunol 2015; 22:389–397 [View Article] [PubMed]
     [Google Scholar]
  16. Mohapatra AK, Mohapatra JK, Pandey LK, Sanyal A, Pattnaik B. Diagnostic potential of recombinant nonstructural protein 3B to detect antibodies induced by foot-and-mouth disease virus infection in bovines. Arch Virol 2014; 159:2359–2369 [View Article] [PubMed]
     [Google Scholar]
  17. Lu Z, Cao Y, Guo J, Qi S, Li D et al. Development and validation of a 3ABC indirect ELISA for differentiation of foot-and-mouth disease virus infected from vaccinated animals. Vet Microbiol 2007; 125:157–169 [View Article] [PubMed]
     [Google Scholar]
  18. Sørensen KJ, de Stricker K, Dyrting KC, Grazioli S, Haas B. Differentiation of foot-and-mouth disease virus infected animals from vaccinated animals using a blocking ELISA based on baculovirus expressed FMDV 3ABC antigen and a 3ABC monoclonal antibody. Arch Virol 2005; 150:805–814 [View Article] [PubMed]
     [Google Scholar]
  19. Sørensen KJ, Madsen KG, Madsen ES, Salt JS, Nqindi J et al. Differentiation of infection from vaccination in foot-and-mouth disease by the detection of antibodies to the non-structural proteins 3D, 3AB and 3ABC in ELISA using antigens expressed in baculovirus. Arch Virol 1998; 143:1461–1476 [View Article] [PubMed]
     [Google Scholar]
  20. Gelkop S, Sobarzo A, Brangel P, Vincke C, Romão E et al. The development and validation of a novel nanobody-based competitive ELISA for the detection of foot and mouth disease 3ABC antibodies in cattle. Front Vet Sci 2018; 5:250 [View Article] [PubMed]
     [Google Scholar]
  21. Hosamani M, Basagoudanavar SH, Tamil Selvan RP, Das V, Ngangom P et al. A multi-species indirect ELISA for detection of non-structural protein 3ABC specific antibodies to foot-and-mouth disease virus. Arch Virol 2015; 160:937–944 [View Article] [PubMed]
     [Google Scholar]
  22. Clavijo A, Zhou E-M, Hole K, Galic B, Kitching P. Development and use of a biotinylated 3ABC recombinant protein in a solid-phase competitive ELISA for the detection of antibodies against foot-and-mouth disease virus. J Virol Methods 2004; 120:217–227 [View Article] [PubMed]
     [Google Scholar]
  23. Malirat V, Neitzert E, Bergmann IE, Maradei E, Beck E. Detection of cattle exposed to foot‐and‐mouth disease virus by means of an indirect ELISA test using bioengineered nonstructural polyprotein 3ABC. Veterinary Quarterly 1998; 20:24–26 [View Article] [PubMed]
     [Google Scholar]
  24. Mohapatra JK, Pandey LK, Sanyal A, Pattnaik B. Recombinant non-structural polyprotein 3AB-based serodiagnostic strategy for FMD surveillance in bovines irrespective of vaccination. J Virol Methods 2011; 177:184–192 [View Article] [PubMed]
     [Google Scholar]
  25. Jaworski JP, Compaired D, Trotta M, Perez M, Trono K et al. Validation of an r3AB1-FMDV-NSP ELISA to distinguish between cattle infected and vaccinated with foot-and-mouth disease virus. J Virol Methods 2011; 178:191–200 [View Article] [PubMed]
     [Google Scholar]
  26. Mahajan S, Mohapatra JK, Pandey LK, Sharma GK, Pattnaik B. Indirect ELISA using recombinant nonstructural protein 3D to detect foot and mouth disease virus infection associated antibodies. Biologicals 2015; 43:47–54 [View Article] [PubMed]
     [Google Scholar]
  27. Clavijo A, Hole K, Li M, Collignon B. Simultaneous detection of antibodies to foot-and-mouth disease non-structural proteins 3ABC, 3D, 3A and 3B by a multiplexed Luminex assay to differentiate infected from vaccinated cattle. Vaccine 2006; 24:1693–1704 [View Article] [PubMed]
     [Google Scholar]
  28. Mackay DK, Forsyth MA, Davies PR, Berlinzani A, Belsham GJ et al. Differentiating infection from vaccination in foot-and-mouth disease using a panel of recombinant, non-structural proteins in ELISA. Vaccine 1998; 16:446–459 [View Article] [PubMed]
     [Google Scholar]
  29. Sørensen KJ, Hansen CM, Madsen ES, Madsen KG. Blocking ELISAs using the FMDV non‐structural proteins 3D, 3AB, and 3ABC produced in the baculovirus expression system. Veterinary Quarterly 1998; 20:17–20 [View Article] [PubMed]
     [Google Scholar]
  30. Inoue T, Parida S, Paton DJ, Linchongsubongkoch W, Mackay D et al. Development and evaluation of an indirect enzyme-linked immunosorbent assay for detection of foot-and-mouth disease virus nonstructural protein antibody using a chemically synthesized 2B peptide as antigen. J Vet Diagn Invest 2006; 18:545–552 [View Article] [PubMed]
     [Google Scholar]
  31. Oem JK, Kye SJ, Lee KN, Park JH, Kim YJ et al. Development of synthetic peptide ELISA based on nonstructural protein 2C of foot and mouth disease virus. J Vet Sci 2005; 6:317 [View Article]
     [Google Scholar]
  32. Gao M, Zhang R, Li M, Li S, Cao Y et al. An ELISA based on the repeated foot-and-mouth disease virus 3B epitope peptide can distinguish infected and vaccinated cattle. Appl Microbiol Biotechnol 2012; 93:1271–1279 [View Article] [PubMed]
     [Google Scholar]
  33. Bergmann IE, Malirat V, Neitzert E, Beck E, Panizzutti N et al. Improvement of a serodiagnostic strategy for foot-and-mouth disease virus surveillance in cattle under systematic vaccination: a combined system of an indirect ELISA-3ABC with an enzyme-linked immunoelectrotransfer blot assay. Arch Virol 2000; 145:473–489 [View Article] [PubMed]
     [Google Scholar]
  34. Grubman MJ, Baxt B. Foot-and-mouth disease. Clin Microbiol Rev 2004; 17:465–493 [View Article] [PubMed]
     [Google Scholar]
  35. O.I.EOIE Standards Commission Foot and mouth disease. In Manual of Standards for Diagnostic Tests and Vaccines, Office International Des Epizooties 2.1.1. Paris, France: 2004b
     [Google Scholar]
  36. Brocchi E, Bergmann IE, Dekker A, Paton DJ, Sammin DJ et al. Comparative evaluation of six ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus. Vaccine 2006; 24:6966–6979 [View Article] [PubMed]
     [Google Scholar]
  37. Chen SP, Ellis TM, Lee MC, Cheng IC, Yang PC et al. Comparison of sensitivity and specificity in three commercial foot-and-mouth disease virus non-structural protein ELISA kits with swine sera in Taiwan. Vet Microbiol 2007; 119:164–172 [View Article] [PubMed]
     [Google Scholar]
  38. Rice JC, Allis CD. Histone methylation versus histone acetylation: new insights into epigenetic regulation. Curr Opin Cell Biol 2001; 13:263–273 [View Article] [PubMed]
     [Google Scholar]
  39. Falk MM, Grigera PR, Bergmann IE, Zibert A, Multhaup G et al. Foot-and-mouth disease virus protease 3C induces specific proteolytic cleavage of host cell histone H3. J Virol 1990; 64:748–756 [View Article] [PubMed]
     [Google Scholar]
  40. Sariya L, Thangthumniyom N, Wajjwalku W, Chumsing W, Ramasoota P et al. Expression of foot and mouth disease virus nonstructural polyprotein 3ABC with inactive 3Cpro in Escherichia coli. Protein Expr Purif 2011; 80:17–21 [View Article] [PubMed]
     [Google Scholar]
  41. Kweon CH, Ko YJ, Kim WI, Lee SY, Nah JJ et al. Development of a foot-and-mouth disease NSP ELISA and its comparison with differential diagnostic methods. Vaccine 2003; 21:1409–1414 [View Article] [PubMed]
     [Google Scholar]
  42. Curry S, Roqué-Rosell N, Zunszain PA, Leatherbarrow RJ. Foot-and-mouth disease virus 3C protease: recent structural and functional insights into an antiviral target. Int J Biochem Cell Biol 2007; 39:1–6 [View Article] [PubMed]
     [Google Scholar]
  43. Sharma GK, Mohapatra JK, Pandey LK, Mahajan S, Mathapati BS et al. Immunodiagnosis of foot-and-mouth disease using mutated recombinant 3ABC polyprotein in a competitive ELISA. J Virol Methods 2012; 185:52–60 [View Article] [PubMed]
     [Google Scholar]
  44. Srisombundit V, Tungthumniyom N, Linchongsubongkoch W, Lekcharoensuk C, Sariya L et al. Development of an inactivated 3Cpro-3ABC (mu3ABC) ELISA to differentiate cattle infected with foot and mouth disease virus from vaccinated cattle. J Virol Methods 2013; 188:161–167 [View Article] [PubMed]
     [Google Scholar]
  45. Zhou Z, Mogensen MM, Powell PP, Curry S, Wileman T. Foot-and-mouth disease virus 3C protease induces fragmentation of the Golgi compartment and blocks intra-Golgi transport. J Virol 2013; 87:11721–11729 [View Article] [PubMed]
     [Google Scholar]
  46. Puckette M, Smith JD, Gabbert L, Schutta C, Barrera J et al. Production of foot-and-mouth disease virus capsid proteins by the TEV protease. J Biotechnol 2018; 275:7–12 [View Article] [PubMed]
     [Google Scholar]
  47. Birtley JR, Knox SR, Jaulent AM, Brick P, Leatherbarrow RJ et al. Crystal structure of foot-and-mouth disease virus 3C protease. New insights into catalytic mechanism and cleavage specificity. J Biol Chem 2005; 280:11520–11527 [View Article] [PubMed]
     [Google Scholar]
  48. Zunszain PA, Knox SR, Sweeney TR, Yang J, Roqué-Rosell N et al. Insights into cleavage specificity from the crystal structure of foot-and-mouth disease virus 3C protease complexed with a peptide substrate. J Mol Biol 2010; 395:375–389 [View Article] [PubMed]
     [Google Scholar]
  49. Herod MR, Gold S, Lasecka-Dykes L, Wright C, Ward JC et al. Genetic economy in picornaviruses: Foot-and-mouth disease virus replication exploits alternative precursor cleavage pathways. PLoS Pathog 2017; 13:e1006666 [View Article] [PubMed]
     [Google Scholar]
  50. Kruger NJ. The Bradford method for protein quantitation. The Protein Protocols Handbook 200917–24
     [Google Scholar]
  51. De Diego M, Brocchi E, Mackay D, De Simone F. The non-structural polyprotein 3ABC of foot-and-mouth disease virus as a diagnostic antigen in ELISA to differentiate infected from vaccinated cattle. Arch Virol 1997; 142:2021–2033 [View Article] [PubMed]
     [Google Scholar]
  52. Waheed U, Parida S, Khan QM, Hussain M, Ebert K et al. Molecular characterisation of foot-and-mouth disease viruses from Pakistan, 2005-2008. Transbound Emerg Dis 2011; 58:166–172 [View Article] [PubMed]
     [Google Scholar]
  53. Ali W, Habib M, Khan RSA, Zia MA, Farooq M et al. Molecular investigation of foot-and-mouth disease virus circulating in Pakistan during 2014-17. Arch Virol 2018; 163:1733–1743 [View Article] [PubMed]
     [Google Scholar]
  54. Shahbaz P, Boz I, Ul Haq S. Adaptation options for small livestock farmers having large ruminants (cattle and buffalo) against climate change in Central Punjab Pakistan. Environ Sci Pollut Res Int 2020; 27:17935–17948 [View Article] [PubMed]
     [Google Scholar]
  55. Klein J, Hussain M, Ahmad M, Afzal M, Alexandersen S. Epidemiology of foot-and-mouth disease in Landhi Dairy Colony, Pakistan, the world largest Buffalo colony. Virol J 2008; 5:1–16 [View Article] [PubMed]
     [Google Scholar]
  56. Nawaz Z, Arshad M, Iqbal Z. Epidemiology of foot and mouth disease in buffaloes and cattle of Punjab using non structural proteins ELISA. Pakistan Journal of Agricultural Sciences 2014; 51:
     [Google Scholar]
  57. Ur-Rehman S, Arshad M, Hussain I, Iqbal Z. Detection and seroprevalence of foot and mouth disease in sheep and goats in Punjab, Pakistan. Transbound Emerg Dis 2014; 61 Suppl 1:25–30 [View Article] [PubMed]
     [Google Scholar]
  58. Farooq U, Irshad H, Ullah A, Latif A, Zahur A et al. Sero-prevalence of foot-and-mouth disease in small ruminants of Pakistan. J Anim Plant Sci 2017; 27:1197–1201
     [Google Scholar]
  59. Chung CJ, Clavijo A, Bounpheng MA, Uddowla S, Sayed A et al. An improved, rapid competitive ELISA using a novel conserved 3B epitope for the detection of serum antibodies to foot-and-mouth disease virus. J Vet Diagn Invest 2018; 30:699–707 [View Article] [PubMed]
     [Google Scholar]
  60. Lee F, Jong M-H, Yang D-W. Presence of antibodies to non-structural proteins of foot-and-mouth disease virus in repeatedly vaccinated cattle. Vet Microbiol 2006; 115:14–20 [View Article] [PubMed]
     [Google Scholar]
  61. Ullah A, Jamal SM, Romey A, Gorna K, Kakar MA et al. Genetic Characterization of Serotypes A and Asia-1 Foot-and-mouth Disease Viruses in Balochistan, Pakistan, in 2011. Transbound Emerg Dis 2017; 64:1569–1578 [View Article] [PubMed]
     [Google Scholar]
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Development of an ELISA to distinguish between foot-and-mouth disease virus infected and vaccinated animals utilising the viral non-structural protein 3ABC
J. Med. Microbiol. 71, 001516 (2022); https://doi.org/10.1099/jmm.0.001516
/content/journal/jmm/10.1099/jmm.0.001516
/content/journal/jmm/10.1099/jmm.0.001516
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