Unconjugated
Monoclonal antibodies (Mabs) against influenza B virus were obtained by immunizing mice with B/Nagasaki/1/87, one of the strains of the B/Victoria group. Immunoprecipitation analysis revealed that individual Mabs precipitated the nucleoprotein (NP), the matrix protein (M) or the hemagglutinin protein (HA). By using these Mabs by the peroxidase-antiperoxidase (PAP) staining method, a rapid detection and identification method for influenza B virus was established. Monolayers of Madin-Darby canine kidney cells in microplates were infected with each-strain and incubated for about 24 h, and then were subjected to the PAP staining method using the Mabs as the first antibody. Influenza B virus strains are classified into two major phylogenetic trees, the B/Victoria group and the B/Yamagata group. When anti-NP and anti-M antibodies were used in the PAP staining method, all 13 influenza B virus strains isolated from clinical specimens between 1940 and 1994 were detected regardless of the antigenic drift of the influenza virus. On the other hand, several anti-HA Mabs which reacted specifically with the strains of the B/Victoria group, did not react with any strain of the B/Yamagata group. In the 1996/97 influenza season in Osaka Prefecture in Japan, two antigenically distinct groups of influenza B virus strains were isolated. They belonged to different phylogenetic trees and were clearly distinguishable by the PAP staining method with anti-HA Mabs.