Detection and Classification of Infectious Bronchitis Viruses Isolated in Korea by Dot-Immunoblotting Assay Using Monoclonal Antibodies

Abstract

Dot-immunoblotting assay (DIA) using five monoclonal antibodies (MAbs) to infectious bronchitis virus (IBV) was used to detect and classify the viruses propagated in embryonated chicken eggs. Using a group-specific MAb 3F5, 10 reference strains and 12 Korean isolates of IBV were successfully detected by DIA, and the lowest virus titer of IBV detected by DIA was approximately less than 10(3.8) mean embryo infective dose/ml. For evaluating the diagnostic efficiency, DIA was compared with the conventional infectious bronchitis (IB) diagnostic method. IBV antigens in allantoic fluid from embryonated eggs inoculated with IB-suspected field samples were specifically detected by DIA within only one or two egg passages, whereas the conventional embryonated egg inoculation method required four to seven egg passages for confirming IBV infection. These results indicated that DIA could significantly reduce time and cost for IB diagnosis. For examining the possibility of classifying IBV by DIA, four strain-specific MAbs, 3A4, 2A3, 6F7, and 2C6, were used. According to the MAb reacting patterns to the IBV antigens, the 10 IBV reference strains were classified into six groups; seven strains belonged to three different groups, and the other three strains each belonged to an individual group. In the case of 12 Korean isolates of IBV, they were classified in six groups. Among the six groups, the MAb reacting patterns of three groups matched those of the IBV reference strains, but the others did not. These data suggest that at least three variant serotypes of IBV exist in Korea.

Development and Application of Reference Antisera against 15 Hemagglutinin Subtypes of Influenza Virus by DNA Vaccination of Chickens

“…As observed in cross-HI tests (Tables 2 and 3), some cross-reactivity was also observed between H7 sera and the H15 antigen in the immunodot blot assay. In previous work, the immunodot blot assay has been used by others for serotyping of the infectious bronchitis virus (14) and has been applied for the detection of influenza virus (11,13). Thus, with more optimization, we also expect that the antisera prepared by DNA vaccines can be used for subtyping of influenza virus using the immunodot blot assay or similar methods.…”

Induction of protective immunity in chickens vaccinated with infectious bronchitis virus S1 glycoprotein expressed by a recombinant baculovirus.

“…Re-identification of the challenge virus was performed using a dotimmunoblotting assay (Song et al, 1998) Protection of chickens with recombinant IBV S1 Protection of chickens with recombinant IBV S1…”

Nephropathogenesis of Chickens Experimentally Infected with Various Strains of Infectious Bronchitis Virus

“…Dot-immunoblotting assay (DIA) using Mab 9B1B6: DIA was conducted as described previously with the following modifications [23]. Two hundred µl of allantoic fluid used for the challenge study was dotted onto 0.45 µm NC membrane (MSI, Westboro, MA) using the 96-well hybridot manifold (BRL, Gaitherburg, MO).…”