Monoclonal-Antibody-Based Immunodot Assay Distinguishes between Viral Hemorrhagic Septicemia Virus (VHSV) and Infectious Hematopoietic Necrosis Virus (IHNV)

Abstract

An immunodot assay has been developed with two monoclonal antibodies that recognize conserved epitopes on the nucleoproteins of viral hemorrhagic septicemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV). Monoclonal antibody 1NDW14D, which recognizes a conserved epitope on the nucleoprotein of IHNV, recognized 80 of 81 IHNV isolates spotted on nitrocellulose, but none of 8 VHSV isolates. Monoclonal antibody IP5B11, which recognizes a conserved epitope on the nucleoprotein of VHSV, reacted with all eight isolates of VHSV, but with none of the IHNV isolates. Neither monoclonal antibody bound to other rhabdoviruses spotted on nitrocellulose: spring viremia of carp virus (SVCV), pike fry rhabdovirus (PFRV), a new Danish eicosid rhabdovirus unrelated to PFRV, and rhabdovirus anguilla (EVX).

The route of entry and progression of infectious haematopoietic necrosis virus in Oncorhynchus mykiss (Walbaum): a sequential immunohistochemical study

“…Nonspecific binding sites were blocked for lh with 5% non-fat powdered milk (Saco Foods, Madison, WI, USA) in PBS. Blocking solution was blotted off and sections were incubated for lh with a anti-nucleocapsid monoclonal antibody (1NDW14D) whieh is specific for a highly conserved region of the nucleoeapsid protein, and has been shown by direct immunofluorescence and immunoblotting to recognize all five IHNV types (Ristow & Arnzen 1989;Ristow, Lorenzen & Vestergaard J0rgensen 1991). Slides were then incubated for 30min with biotinylated goat anti-mouse polyelonal antibody, followed by a lh incubation with phosphatase-conjugated avidin-biotin complex (Vectastain-ABC mouse IgG kit, Vector Laboratories, Burlingame, CA, USA).…”

Development and Evaluation of a Blocking Enzyme-Linked Immunosorbent Assay and Virus Neutralization Assay To Detect Antibodies to Viral Hemorrhagic Septicemia Virus

“…We demonstrated by Western blotting that the nucleocapsid monoclonal antibody binding was specific to VHSV versus SVCV, another rhabdovirus that is present in Wisconsin. Previous studies showed a lack of cross-reaction between the antibody used herein and spring viremia of carp virus, infectious hematopoietic necrosis virus, pike fry rhabdovirus, or rhabdovirus anguilla (26,56). According to the manufacturer of the antibody (Aquatic Diagnostics, Stirling, Scotland), no cross-reaction of the antibody occurs with nodavirus, infectious salmon anemia virus, koi herpesvirus, salmon alphavirus (1, 2, and 3), or Piscirickettsia salmonis infected cells.…”

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

“…Most probably, broad antigenic variations exist regarding virion configuration (Luo et al, 1990;Ristow, 1991), therefore neutralisation mechanisms have not been clarified yet. Interestingly, MAb 2E1 did not react in Western blotting even after separation with non-reducing conditions.…”

“…The ELISA method can only demonstrate the presence of viral immunogenic particles, and not infectivity of the virus, therefore, no significant correlation was attributed to infectious titre of particular isolates and the ELISA test, due to differences existing in relative amounts of incomplete or decomposed viral particles (Ristow, 1991).…”

“…The methods suitable for epidemiological tracing aiming at determination of the origin of infections are also required -isolation of the virus in susceptible cell lines, immunoassays such as immunofluorescence, ELISA systems and virus neutralisation and/or gene amplification asays (Rodak et al, 1993;Einer-Jensen et al, 2002). Regarding the fact that rhabdoviral glycoproteins can in most cases induce production of neutralizing antibodies in the infected host, as confirmed by data from the literature (Volk et al, 1982;Lorenzen et al, 1990;Luo et al, 1990;Ristow, 1991), there is the possibility of preparing a successful vaccine to SVC virus, i.e. recombinant subunitary vaccine.…”