Rapid Fluorescent Antibody Tests for Infectious Hematopoeitic Necrosis Virus (IHNV) Utilizing Monoclonal Antibodies to the Nucleoprotein and Glycoprotein

Abstract

A quick direct fluorescent antibody test incorporating a monoclonal antibody, 1NDW14D, reacting with the nucleoprotein of infectious hematopoietic necrosis virus (IHNV), revealed the presence of the 1989 Dworshak isolates of the virus 8 h after Chinook salmon embryo (CHSE-214) cells were incubated with fish homogenates containing the virus. An indirect fluorescence assay was performed to compare the ability of the anti-nucleoprotein antibody, 1NDW14D, with that of an anti-glycoprotein antibody, 3GH135L, to detect IHNV. Although the nucleoprotein of IHNV is synthesized before the glycoprotein, both antibodies detected the presence of virusinfected cells with equal efficiency when tested on several isolates of IHNV. Both antibodies stained caps of glycoprotein or nucleoprotein in the cytoplasm contiguous with the plasma membrane of infected cells within 6-8 h after infection.

Infectious hematopoietic necrosis virus: advances in diagnosis and vaccine development

“…Nevertheless, VNT is still being used to detect IHNV infection without sacrificing the fish (Jenčič et al, 2014). More rapid tests based on viral antigen recognition, such as the direct and indirect fluorescent antibody tests (FAT/IFAT) (Arnzen et al, 1991; LaPatra et al, 1989; Woo, Leatherland & Bruno, 2011), ELISA (Adams & Thompson, 2011; Kim et al, 2008), peroxidase immunohistochemical and alkaline phosphatase immunocytochemical (APIC) staining (Drolet, Rohovec & Leong, 1993; Yamamoto et al, 1990), and western blotting (Ristow et al, 1993) have been successfully developed. FAT/IFAT and APIC staining are often used to detect the presence of IHNV in infected fishes through immunostaining of tissue imprints or fixed tissue sections.…”

Peer Review #2 of "Infectious hematopoietic necrosis virus: advances in diagnosis and vaccine development (v0.1)"

“…Nevertheless, VNT is still being used to detect IHNV infection without sacrificing the fish (Jenčič et al 2014). More rapid tests based on viral antigen recognition, such as the direct and indirect fluorescent antibody tests (FAT/IFAT) (Arnzen et al 1991;Lapatra et al 1989;Woo et al 2011), ELISA (Adams & Thompson 2011;Kim et al 2008), peroxidase immunohistochemical and alkaline phosphatase immunocytochemical (APIC) staining (Drolet et al 1993;Yamamoto et al 1990), and western blotting (Ristow et al 1993) have been successfully developed. FAT/IFAT and APIC staining are often used to detect the presence of IHNV in infected fishes through immunostaining of tissue imprints or fixed tissue sections.…”

Trout and Salmon Viruses

“…The system was sensitive for five known IHNV types and no cross-reactivity was noted with six other fish rhabdoviruses or with IPNV. According to Arnzen et al (1991) the fluorescent antibody technique (FAT) is as sensitive as plaque assay and requires less time to obtain a confirmed diagnosis; however, monoclonal antibody against IHNV nucleoprotein and glycoprotein detected stages of virus replication in cell cultures as early as 6-8 hours postinoculation. LaPatra et al (1989b) found that polyclonal and monoclonal antibody can be used to detect IHNV in blood and organ smears of juvenile salmonids and in ovarian fluid of adults.…”