A 12-residue epitope displayed on phage T7 reacts strongly with antibodies against foot-and-mouth disease virus
Foot-and-mouth disease (FMD) is a major threat to the livestock industry worldwide. Despite constant surveillance and effective vaccination, the perpetual mutations of the foot-and-mouth disease virus (FMDV) pose a huge challenge to FMD diagnosis. The immunodominant region of the FMDV VP1 protein (residues 131–170) displayed on phage T7 has been used to detect anti-FMDV in bovine sera. In the present study, the functional epitope was further delineated using amino acid sequence alignment, homology modelling and phage display. Two highly conserved regions (VP1145–152 and VP1159–170) were identified among different FMDV serotypes. The coding regions of these two epitopes were fused separately to the T7 genome and displayed on the phage particles. Interestingly, chimeric phage displaying the VP1159–170 epitope demonstrated a higher antigenicity than that displaying the VP1131–170 epitope. By contrast, phage T7 displaying the VP1145–152 epitope did not react significantly with the anti-FMDV antibodies in vaccinated bovine sera. This study has successfully identified a smaller functional epitope, VP1159–170, located at the C-terminal end of the structural VP1 protein. The phage T7 displaying this shorter epitope is a promising diagnostic reagent to detect anti-FMDV antibodies in vaccinated animals.
KeywordsDiagnostic reagent ELISA Foot-and-mouth disease Phage display VP1 epitope
The authors thank Dr. Baljit Singh (Universiti Putra Malaysia) for technical assistance in collection of bovine sera and J.Y. Chia for technical assistance in homology modelling.
Compliance with ethical standards
Conflict of interest
Wong CL declares that she has no conflict of interest. Yong CY declares that he has no conflict of interest. Muhamad A declares that she has no conflict of interest. Syahir A declares that he has no conflict of interest. Omar AR declares that he has no conflict of interest. Sieo CC declares that she has no conflict of interest. Tan WS declares that he has no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Adams MH (1959) Assay of phages by the agar layer method. In: Adams MH (ed) Bacteriophages, 1st edn. Interscience Publishers, New York, pp 450–451Google Scholar
- Biasani M, Bienert S, Waterhouse A, Arnold K, Studer G, Schmidt T, Kiefer F, Gallo Cassarino T, Bertoni M, Bordoli L, Schwede T (2014) SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Res 42:252–258. https://doi.org/10.1093/nar/gku340 CrossRefGoogle Scholar
- Carrillo C (2012) Foot-and-mouth disease virus genome. In: Garcia ML, Romanowski V (eds) Viral genomes-molecular structure, diversity, gene expression mechanisms and host-virus interactions, 1st edn. InTech, Rijeka, pp 53–68Google Scholar
- Colling A, Morrissy C, Barr J, Meehan G, Wright L, Goff W, Gleeson LJ, van der Heide B, Riddell S, Yu M, Eagles D, Lunt R, Khounsy S, Than Long NG, Phong Vu P, Than Phuong N, Tung N, Linchongsubongkoch W, Hammond J, Johnson M, Johnson WO, Unger H, Daniels P, Crowther JR (2014) Development and validation of a 3ABC antibody ELISA in Australia for foot and mouth disease. Aust Vet J 92:192–199. https://doi.org/10.1111/avj.12190 CrossRefPubMedGoogle Scholar
- Crowther JR, Farias S, Carpenter WC, Samuel AR (1993) Identification of a fifth neutralizable site on type O foot-and-mouth disease virus following characterization of a single and quintuple monoclonal antibody escape mutants. J Gen Virol 74:1547–1553. https://doi.org/10.1099/0022-1317-74-8-1547 CrossRefPubMedGoogle Scholar
- Curry S, Fry E, Blakemore W, Abu-Ghazaleh R, Jackson T, King A, Lea S, Newman J, Rowlands D, Stuart D (1996) Perturbations in the surface structure of A22 Iraq foot-and-mouth disease virus accompanying coupled changes in host cell specificity and antigenicity. Structure 4:135–145. https://doi.org/10.1016/S0969-2126(96)00017-2 CrossRefPubMedGoogle Scholar
- Fowler VL, Bashiruddin JB, Maree FF, Mutowembwa P, Bankowski B, Gibson D, Cox S, Knowles N, Barnett PV (2011) Foot-and-mouth disease marker vaccine: cattle protection with a partial VP1 G-H loop deleted virus antigen. Vaccine 29:8405–8411. https://doi.org/10.1016/j.vaccine.2011.08.035 CrossRefPubMedGoogle Scholar
- Kitson JDA, McCahon D, Belsham GJ (1990) Sequence analysis of monoclonal antibody resistant mutants of type O foot-and-mouth disease virus: evidence for involvement of the three surface exposed capsid proteins in four antigenic sites. Virology 179:26–34. https://doi.org/10.1016/0042-6822(90)90269-W CrossRefPubMedGoogle Scholar
- Netter HJ, Macnaughton TB, Woo WP, Tindle R, Gowans EJ (2001) Antigenicity and immunogenicity of novel chimeric hepatitis B surface antigen particles with exposed hepatitis C virus epitopes. J Virol 75:2130–2141. https://doi.org/10.1128/JVI.75.5.2130-2141.2001 CrossRefPubMedPubMedCentralGoogle Scholar
- OIE (2011) Foot and mouth disease. Terrestrial Animal Health Code. http://www.oie.int/eng/A_FMD2012/docs/en_chapitre_1.8.5.pdf. Accessed 15 Aug 2017
- OIE (2017) Foot and mouth disease (Infection with foot and mouth disease virus). Terrestrial Animal Manual. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.01.08_FMD.pdf. Accessed 15 Aug 2017
- OIE, FAO (2012) The Global Foot and Mouth Disease Control Strategy—strengthening animal health systems through improved control of major diseases. http://www.oie.int/doc/ged/D11886.PDF. Accessed 2 Feb 2018
- Ptaff E, Thiel HJ, Strohmaier K, Schaller H (1988) Analysis of neutralizing epitopes on foot-and-mouth disease virus. J Gen Virol 62:2033–2020Google Scholar
- Rae BP, Elliott RM (1986) Characterization of the mutations responsible for the electrophoretic mobility differences in the NS proteins of vesicular stomatitis virus New Jersey complementation group E mutants. J Gen Virol 67:2635–2643. https://doi.org/10.1099/0022-1317-67-12-2635 CrossRefPubMedGoogle Scholar
- Robinson TP, Thornton PK, Franceschini G, Kruska RL, Chiozza F, Notenbaert A, Cecchi G, Herrero M, Epprecht M, Fritz S, You L, Conchedda G, See L (2011) Global livestock production systems. Rome, FAO of the United Nations and ILRI, p 152. http://www.fao.org/docrep/014/i2414e/i2414e.pdf. Accessed 19 Aug 2017
- Shi Y, Mowery RA, Ashley J, Hentz M, Ramirez AJ, Bilgicer B, Slunt-Brown H, Borchelt DR, Shaw BF (2012) Abnormal SDS-PAGE migration of cytosolic proteins can identify domains and mechanisms that control surfactant binding. Protein Sci 21:1197–1209. https://doi.org/10.1002/pro.2107 CrossRefPubMedPubMedCentralGoogle Scholar
- Uttenthal A, Parida S, Rasmussen TB, Paton DJ, Haas B, Dundon WG (2010) Strategies for differentiating infection in vaccinated animals (DIVA) for foot-and-mouth disease, classical swine fever and avian influenza. Expert Rev Vaccines 9:73–87. https://doi.org/10.1586/erv.09.130 CrossRefPubMedGoogle Scholar