Detection of spring viraemia of carp virus (SVCV) with monoclonal antibodies

Abstract

Monoclonal antibodies (MAbs) against spring viraemia of carp virus (SVCV), a severe disease in cyprinid fish, were prepared. Nine MAbs were characterised using Western blotting (WB) where all reacted with glycoprotein G, except for MAb 2E1, which showed no reactivity in WB. All nine MAbs showed specificity in an immunoperoxidase test. In ELISA assays, their titres ranged between 1:32 000 and 1:128 000. A panel of SVCV isolates from different European regions were set up and examined by sandwich ELISA assay using the MAbs at a concentration of 15 µg/ml. Only MAb 4C12/3C8 showed low sensitivity in most of the isolates at an absorbance of A 450 . Of the other MAbs, even the lowest absorbance value measured exceeded cut-off for evaluation of the whole reaction. No cross-reaction with the infectious haematopoietic necrosis virus (IHNV), viral haemorrhagic septicaemia virus (VHSV) or infectious pancreatic necrosis virus (IPNV) was demonstrated. 2E1 did not show cross-reactivity with PFRV classified in genogroup III-IV and reacted with a Czech SVCV isolate; its identity was confirmed by means of RT PCR assay. The others MAbs reacted positively with PFRV F4 reference strain, isolated from Esox lucius L. (genogroup III).

Development and characterization of monoclonal antibodies to spring viraemia of carp virus

“…Monoclonal antibodies (mAbs), as useful tools, have been used widely for development of effective diagnosis of fish viral pathogens, such as infectious hematopoietic necrosis virus (IHNV; Schultz et al, 1985), infectious pancreatic necrosis virus (IPNV; Wolski et al, 1986), viral hemorrhagic septicemia virus (VHSV; Sanz et al, 1992), epizootic hematopoietic necrosis virus (EHNV; Monini and Ruggeri, 2002), and so on. Concerning SVCV, mAbs specific against the SVCV had been recently developed and applied to detect the viral antigens by sandwich ELISA (Bing et al, 2007;Reschova et al, 2007). However, problems with mAbs specificity still need to be resolved (Dixon and Longshaw, 2005;Reschova et al, 2007), and precise reports on immunoassays such as virus neutralization assay, immunodot blotting and immunohistochemistry assay with mAbs against SVCV were rare.…”

“…Concerning SVCV, mAbs specific against the SVCV had been recently developed and applied to detect the viral antigens by sandwich ELISA (Bing et al, 2007;Reschova et al, 2007). However, problems with mAbs specificity still need to be resolved (Dixon and Longshaw, 2005;Reschova et al, 2007), and precise reports on immunoassays such as virus neutralization assay, immunodot blotting and immunohistochemistry assay with mAbs against SVCV were rare. Furthermore, neutralizing mAbs against SVCV have yet not been reported.…”

Selection and characterization of single-chain recombinant antibodies against spring viraemia of carp virus from mouse phage display library

“…Cross-reactivity with several other aquatic viruses was tested with the scFv antibodies. Especially, SVCV and PFRV share antigenic determinants with G, N and M proteins and their differentiation by means of serologic tests using polyclonal or monoclonal antibodies is often problematic (Reschova et al, 2007). In the present study, the isolated eight clones (P2H12, P2G6, P1A5, P1F7, P1B7, P1H2, P1E7 and P2F1) which recognized SVCV specifically, and had no cross-reactivity with PFRV, STIV, VNNV, VHSV and IHNV.…”

“…The produced 29 kDa specific scFv antibodies were confirmed by Western blot assay. Monoclonal antibodies have been used in protocols developed for the rapid and sensitive detection and diagnosis of SVCV, for example, antigen capture ELISA, sandwich ELISA and IFA (Dixon and Longshaw, 2005;Reschova et al, 2007). In order to further characterization of scFvs in this study, they were applied to assess specificity by ELISA and immunofluorescent.…”

Generation and characterization of monoclonal antibodies against the flounder Paralichthys olivaceus rhabdovirus

“…Similar results were also reported in studies developing MAbs agaist a grouper iridovirus (Shi et al, 2003), haemocytes of scallop (Xing and Zhan, 2005), lymphocystis disease virus (Cheng et al, 2006), S. maximus rhabdovirus (Zhou et al, 2006) in aquaculture research, respectively. Concerning fish rhabdoviruses, previous reports on MAbs focused mainly on virus detection using the antigen capture ELISA, sandwich ELISA and immunoperoxidase techniques (Schultz et al, 1985;Mourton et al, 1992;Sanz et al, 1992;Reno and Lannan, 1992;Reschova et al, 2007). With the development of modern immunological techniques, many new diagnosis and characterization methods of virus pathogens have been established in virtue of MAbs.…”