Detection of Rinderpest Virus Using N-Protein Monoclonal Antibodies R.A.Shah 1 ,M.C.Joseph,G.Butchaiah 2 ,M.Malik*,R.K.SinghandC.S.Bakshi National Biotechnology Centre, Indian Veterinary Research Institute, Izatnagar 243 122, UP, India; 1 Sher-e-Kashmir University of Science and Technology, Srinagar, J&K, India; 2 Rajiv Gandhi College of Veterinary and Animal Sciences, Pondicherry, India *Correspondence: E-mail: meenumalik@yahoo.com Shah, R.A., Joseph, M.C., Butchaiah, G., Malik, M., Singh, R.K. and Bakshi, C.S., 2004. Detection of rinderpestvirususingN-proteinmonoclonalantibodies. Tropical Animal Health and Production, 36(1),11^ 25 ABSTRACT Apanelofmonoclonalantibodies(mAbs)wasgeneratedagainsttheRBOKstrainofrinderpestvirus (RPV).AllofthemboundtotheNproteinofRPV.TheantigencaptureELISAusingthemAbscould detect the virus in crude viral preparations. The mAb 12BF8.1.1 showed higher reactivity with cell- associated (CA) virus, whereas the mAbs 12AD10.1.1, 12BD7.1.1 and 12DG7.1.1 showed higher reactivity with extracellular virus (hereafter referred to as cell-free (CF) virus).The mAbs 12BF8.1.1 and12AD10.1.1coulddetectthevirusininfectedVerocellculturesupernatants(CCS)asearlyas24h post-cytopathic e¡ect (CPE) initiation. Detergent treatment (Triton X-100) of RPV preparations enhanced the binding of the mAbs to the virus. All the seven mAbs showed speci¢c £uorescence in virus-infectedcellcultures.Theimmuno£uorescence(IFA)usingmAbswasfoundtobemoresensitive andreliablethantheimmunoperoxidasetest(IPT)fordetectionofrinderpest. Keywords: rinderpestvirus,monoclonalantibodies,N-protein,virusdetection,ELISA,immuno£uor- escenceassay,immunoperoxidasetest Abbreviations: CA,cell-associated;CCS,crudecellculturesupernatant;CDV,caninedistempervirus; CF,cell-free;CPE,cytopathice¡ect;DGP,densitygradientpuri¢ed;ELISA,enzyme-linkedimmuno- sorbent assay; IFA, indirect £uorescence assay; IPT, immunoperoxidase test; mAb, monoclonal antibody; MHIS, mouse hyperimmune serum; MV, measles virus; PPRV, peste des petits ruminants virus; RHIS, rabbit hyperimmune serum; RIPA, radioimmunoprecipitation assay; RPV, rinderpest virus INTRODUCTION Rinderpestisaneconomicallyimportant,highlycontagiousandlethalvirusdiseaseof cloven-footedanimals,particularlycattleandbu¡aloes.Rinderpestvirus(RPV)along withmeaslesvirus(MV),caninedistempervirus(CDV)and peste des petits ruminants virus(PPRV)belongstothemorbillivirusgenusinthefamily Paramyxoviridae (Gibbs et al., 1979). All morbilliviruses are antigenically closely related, as evidencedby the occurrenceofcross-reactionsusingpolyvalentserum.Monoclonalantibodies(mAbs) have been produced and used to antigenically characterize MV, CDV and RPV (Norrby et al., 1985; Orwell et al., 1985; Sato et al., 1985; Bhavani et al., 1989; Tropical Animal Health and Production,36(2004)11^25 # 2004KluwerAcademicPublishers.PrintedintheNetherlands 11