Diagnostic Potential of Polyclonal Antibodies Against Bacterially Expressed Recombinant Coat Protein of Alfalfa mosaic virus

Abstract

Khatabi, B., He, B., and Hajimorad, M. R. 2012. Diagnostic potential of polycional antibodies against bacteriaily expressed recombinant coat protein of Alfalfa mosaic virus. Plant Dis. 96; 1352-1357.Alfalfa mosaic virus (AMV), a pathogen of a wide range of plant species, including Glycine max (soybean), is poorly immunogenic. Polycional antibodies were generated against bacteriaily expressed recombinant coat proteins (rCPs) of two biologically distinct AMV strains in rabbits and compared with those raised against native and glutaraldehyde-treated virions of the same strains. Analyses showed that sera against rCPs had comparable antibody titers in indirect enzyme-linked immunosorbent assay with those raised against virions when soybean sap containing bomologous viruses served as antigens. Polycional antibodies against rCPs were specific, sensitive, and detected all AMV isolates that originated from soybean fields from geographically different regions of the United States. Comparison of CP genes of these isolates showed 96 to 99 and 96 to 100% nueleotide and amino acid sequence identities, respectively, suggesting that they are all closely related. This was further confirmed by phylogenetic analysis where they were all clustered together along with representative AMV strains belonging to group I. Collectively, our data demonstrate that, despite poor immunogenicity of AMV, polycional antibodies against rCP are effective probes for detection and diagnosis ofthe virus.Alfalfa mosaic virus (AMV), the type species of the genus Alfamovirus within the family Bromoviridae, has a broad host range infecting more than 400 plant species in over 51 dicotyledonous families woridwide (7,23), AMV exhibits considerable variability in pathogenieity, transmissibility, and symptom expression (12,23). AMV is an emerging viral problem of soybean {Gly-cine max (L.) Merrill)

Production of a Polyclonal Antibody against the Recombinant Coat Protein of the Sugarcane Mosaic Virus and Its Application in the Immunodiagnostic of Sugarcane

“…SDS-PAGE analysis showed that the expression of the recombinant SCMVCp fusion protein was found to be plentiful in the total extract and in the insoluble fraction of the pellet debris of E. coli, with a molecular size of around 44 kDa, but was not presented in the supernatant as a soluble protein fraction (Figure 2A). This result is consistent with the experiments previously reported that the recombinant virus coat protein was expressed in the insoluble protein fraction [13,15]. Interestingly, the expression of the recombinant SCMVCp protein was not dose-dependent on the inducer IPTG.…”

“…To overcome this problem, the serological method using a polyclonal antibody against a recombinant coat protein, was commonly used in immunodiagnostics [11]. Several studies have reported on the production of polyclonal antibodies against recombinant proteins, such as the antibodies against coat protein of Pelargonium zonate spot virus [13], cucumber mosaic virus [14], or alfalfa mosaic virus [15].…”

Expression and Production of Polyclonal Antibodies against Recombinant Coat Protein of Peanut bud necrosis virus

“…Banana streak MY virus (BSMYV) antiserum (1:2000) was used in antigen coated plate-enzyme linked immunosorbent assay (ACP-ELISA) for the detection of virus in banana plants [27]. Alfalfa mosaic virus (AMV) polyclonal antiserum was used for the detection of virus in soybean plants in USA [28]. Our results with CP are in agreement with the results obtained for antibodies raised against recombinant Watermelon bud necrosis virus [29], Potato mop-top virus [30], Potato virus Y [31], Potato virus X [31].…”

The Comparison of Antibodies Raised Against PLRV with Two Different Approaches - Viral Particles Purification and Recombinant Production of CP

“…Therefore, polyclonal antibodies raised against purified virions contaminated with host tissue components, cross react with host antigens and often give variable background reactions, thus limiting their use in ELISA based diagnostic methods. Recombinant polyclonal antibodies (Pab) specific to Tomato spotted wilt virus (TSWV) [11], Potato virus Y [12], Alfalfa mosaic virus (AMV) Khatabi et al [13] have been generated and used for serological detection of cited viruses [14]. centrifuged at 12.000 rpm for 15 min.…”