Fig. 4

The detection of BoAHV1 IgG levels in cattle serum. (A) The serum samples, originally labeled as 72, 189,119, and 200,178, provided by a local dairy farmer, were tested using the AsurDx ™ Infectious Bovine Rhinotracheitis (IBR) gB Antibody ELISA Test Kit (BioStone Animal Health, Southlake, TX, USA, cat# 10074-05). The positive and negative antibody controls were included with the ELISA kit. (B-F) Purified virions and mock-infected cell lysates of MDBK cells (MICL) of approximately 3.5 µg of total proteins were dot-loaded onto NC membranes. After blocking with 5% non-fat milk in PBST, the membranes were incubated with sera 72 (1:3000), 189,119 (1:3000), 200,178 (1:3000), as well as the positive (1:1000) and negative (1:1000) antibody controls included with the ELISA kit, respectively, followed by incubation with rabbit anti-bovine IgG-HRP (1:8000). The immune reaction was developed using Clarity Western ECL substrate. The images shown are representative of three independent experiments