Development of a quick dot blot assay for the titering of bovine ephemeral fever virus

• Published in: BMC veterinary research 2019-09, Vol.15 (1), p.313-313, Article 313
• Main Authors: Cheng, Li-Ting, Zeng, Yu-Jing, Chu, Chun-Yen, Wang, Hsian-Yu
• Format: Article
• Language: English
• Subjects: Animal vaccines; Animals; Antibodies; Antibodies, Viral; Antigens; antiserum; Assaying; Biomedical laboratory equipment; Bovidae; Bovine ephemeral fever; Bovine ephemeral fever virus; Cattle; Cattle diseases; Cattle industry; Cell culture; Cell Line; Cricetinae; Dairy cattle; dairy farming; Dairy farms; Diagnosis; Dot blot assay; E coli; Economic aspects; Economic impact; Ephemeral Fever - virology; Ephemeral Fever Virus, Bovine - isolation & purification; Escherichia coli; Farms; Fever; G protein; G proteins; Gene Expression Regulation, Viral; Identification and classification; Immunoblotting; Immunoblotting - methods; Immunoblotting - veterinary; Immunoglobulins; inactivated vaccines; manufacturing; Methodology; Methods; Muscles; N protein; Noise; Particulates; Plaque assay; Production management; Proteins; Rabbits; Signal to noise ratio; Sucrose; Vaccines; Vector-borne diseases; Veterinary medicine; viral antigens; viral load; Viral proteins; Viral Proteins - genetics; Viral Proteins - metabolism; Viruses
• ISSN: 1746-6148; 1746-6148
• DOI: 10.1186/s12917-019-2059-6

Bovine ephemeral fever virus (BEFV) causes fever and muscle stiffness in cattle, resulting in negative economic impact for cattle and dairy farms. During the manufacturing process of inactivated vaccine for virus control, it is important to determine the virus titer, but traditional methods such as plaque assay and TCID assay require days of waiting time. We sought to develop a quick dot blot assay for BEFV titering. Three different kinds of BEFV antigens were prepared to raise primary antibodies for BEFV detection in dot blot assays: 1) purified BEFV particles, 2) Escherichia coli (E. coli)-expressed BEFV G1 region, and 3) E. coli-expressed BEFV N protein. Results showed that antibodies raised against purified BEFV particles can detect BEFV particles, but it also showed a high background level from the proteins of BHK-21 cells. Antibodies raised against E.coli-expressed BEFV G1 region could not detect BEFV particles in dot blot assays. Finally, antibodies raised against E.coli-expressed BEFV N protein detected BEFV particles with a high signal-to-noise ratio in dot blot assays. E.coli-expressed N protein is a suitable antigen for the production of antiserum that can detect BEFV particles with a high signal-to-noise ratio. A dot blot assay kit using this antiserum can be developed as a quick and economical way for BEFV titering.