Development of a Monoclonal Antibody-based Immunohistochemical Method for Detecting Newcastle Disease Virus

Abstract

ND Virus Detection in IHC birds is responsible for outbreaks of ND (Heckert et al. 1996, Kinde et al. 2005). Therefore, investigation of the pathogenesis of ND in poultry and wild birds is necessary to minimize the risk of viral transmission between wild birds and poultry. Immunohistochemical (IHC) analysis, which is performed to detect the antigen on histological sections of formalin-fixed, paraffin-embedded tissues, can be a useful research tool for elucidating the pathogenesis of infectious diseases. However, IHC analysis for detecting ND virus is not commonly performed worldwide, probably because of the lack of commercially available suitable primary antibodies. Establishing an IHC method for ND virus detection would facilitate the progress in this research field. In this study, we developed a monoclonal antibody (mAb) applicable to IHC analysis for ND virus detection.

Susceptibility and Pathogenesis of Eurasian Tree Sparrows Experimentally Inoculated with Velogenic Newcastle Disease Virus

“…Five birds (nos. [16][17][18][19][20] in Group B exhibited HI antibody titers of 1:16, ,1:4, 1:256, 1:64, and 1:8, respectively. A nonparametric Kruskal-Wallis test showed a significant difference in antibody levels among the three groups (P ¼ 0.002).…”

“…Immunohistochemistry (IHC) was performed to detect ND viral antigens on histologic sections, as reported previously (19). A mouse anti-NDV nucleoprotein monoclonal antibody (clone 22C483, dilution 1:3200) was used as the primary antibody.…”

“…Virus. The velogenic NDV strain APMV1/chicken/Japan/Fukuoka-1/2004 (JP/Fukuoka-1/2004) was used in this study (18). The virus was isolated from diseased broilers in Japan in 2004 and is classified as genotype VII (18).…”

“…The velogenic NDV strain APMV1/chicken/Japan/Fukuoka-1/2004 (JP/Fukuoka-1/2004) was used in this study (18). The virus was isolated from diseased broilers in Japan in 2004 and is classified as genotype VII (18). The stock virus was propagated for 2-3 days in the allantoic cavity of 9-day-old embryonated chicken eggs at 37 C. Fresh infectious allantoic fluid was harvested and stored at À80 C until use.…”

Pathological and virological analysis of concurrent disease of chicken anemia virus infection and infectious bronchitis in Japanese native chicks

“…IHC was performed to detect avian viruses. The primary antibodies used were as follows: mouse monoclonal antibodies to CAV (clone MoCAV/E6, 1:8,000 dilution, laboratory-made) [33], IBV (clone IB95, 1:5,000 dilution, Hytest Corp., Turku, Finland), infectious bursal disease virus (IBDV) (clone IBD105, 1:2,000 dilution, Hytest Corp.), type A influenza virus (AIV) (clone GA2B, 1:500 dilution, Abcam Corp, Cambridge, U.K.), and Newcastle disease virus (NDV) (clone 22C483, 1:12,800 dilution, laboratory-made) [35] and rabbit immune serum against fowl adenovirus (FAV) (1:5,000 dilution, laboratory-made) [22].…”