Date: 06 Sep 2012
Expression of Foot-and-Mouth Disease Virus Non-Structural Protein, 3D in Insect Cells and its Application in Detection of Anti-FMDV Antibodies
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Non-structural proteins (NSPs) based diagnostics are useful for large-scale sero-surveillance of foot-and-mouth disease (FMD) and to monitor viral activity as a follow up to the vaccination campaign in FMD endemic countries like India which aim at disease control through vaccination. These diagnostics are also handy in the serology of import/export of cloven-footed animals. In the present study, non-structural protein RNA polymerase (3D gene) of FMD virus (FMDV) was expressed using baculovirus expression system. Protein expression was analyzed by SDS-PAGE and confirmed by its immuno-reactivity with serum from a FMDV infected bovine, in the western blot. Recombinant 3D protein was purified and evaluated in the indirect ELISA with 1072 cattle serum samples. Diagnostic sensitivity and specificity of the assay were found to be 92 and 100 %, respectively, when tested with cattle sera of known FMD status. The 3D based ELISA developed here is useful for screening the animals as an adjunct to other NSP based diagnostics available for routine serosurveillance of FMD.
Alonso Fernandez A, Auge de Melo P, Gomes I, Rosemberg F. The use of virus infection associated antigen (VIA) on the detection of cattle exposed to FMDV. Bol Cent Panam Fiebre Aftosa. 1975;17–18:17–22.
Alonso A, Gomes MPD, Martins MA, Sondahl MS. Detection of foot-and-mouth disease virus infection-associated antigen antibodies: comparison of the enzyme-linked immunosorbent assay and agar gel immunodiffusion tests. Prev Vet Med. 1990;9:233–40.CrossRef
Anon (2004) Manual of standards for diagnostic tests and vaccines. Office Internationale des Epizooties (6th edition). Office Internationale des Epizooties, Paris, 1, p. 190–216.
Bergmann IE, Augé de Mello P, Neitzert E, Beck E, Gomes I. Diagnosis of persistent aphthovirus infection and its differentiation from vaccination response in cattle by use of enzyme-linked immunoelectrotransfer blot analysis with bioengineered nonstructural viral antigens. Am J Vet Res. 1993;6:825–31.
Bergmann I, Malirat V, Neitzert E, Beck E, Panizzutti N, Sanchez C. 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;145:473–9.PubMedCrossRef
Cowan KM, Graves JG. A third antigenic component associated with foot-and-mouth disease virus infection. Virol. 1996;30:528–40.
Gebauer F, La Torre JC, Gomes I, Mateu MG, Barahona H, Tiraboschi B, Bergmann I, de Mello PA, Domingo E. Rapid selection of genetic and antigenic variants of FMDV during persistence in cattle. J Virol. 1988;62:2041–9.PubMed
Hoss A, Moarefi I, Scheidtmann KH, Cisek LJ, Corden JL, Dornreiter I, Arthur AK, Fanning E. Altered phosphorylation pattern of simian virus 40 T antigen expressed in insect cells by using a baculovirus vector. J Virol. 1990;64:4799–807.PubMed
Neitzert E, Beck E, Augé de Mello P, Gomes I, Bergmann IE. Expression of the aphthovirus RNA polymerase gene in E. coli and its use together with other bioengineered non-structural antigens in detection of late persistent infections. Virology 1991; 184:799–804
O’Donnell VK, Smitsaart E, Cetra B, Duffy S, Finelli J, Boyle D, Draghi G, Fondevila N, Schudel AA. Detection of virus infection-associated antigen and 3D antibodies in cattle vaccinated against foot-and-mouth disease. Rev Sci Tech Off Int Epiz. 1997;16:833–40.
Pinto AA, Garland AJM. Immune response to virus-infection-associated (VIA) antigen in cattle repeatedly vaccinated with foot-and-mouth disease virus inactivated by formalin or acetylethyleneimine. J Hyg. 1979;82:41–50.CrossRef
Uttenthal A, Parida S, Rasmussen TB, Paton DJ, Haas B, Dundon WG. Strategies for differentiating infection in vaccinated animals (DIVA) for foot-and-mouth disease, classical swine fever and avian influenza. Expert Rev Vac. 2010;9:73–87.CrossRef
Sorenson KJ, Madsen KG, Madsen ES, Salt JS, Nqindi J, Mackay DKJ. 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.
- Expression of Foot-and-Mouth Disease Virus Non-Structural Protein, 3D in Insect Cells and its Application in Detection of Anti-FMDV Antibodies
Indian Journal of Virology
Volume 23, Issue 3 , pp 326-332
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- Foot-and-mouth disease virus
- Non-structural protein