Binary Ethylenimine Inactivated Japanese Encephalitis Virus Antigen Reveals Hemagglutination英文文献资料.docVIP

Open Journal of Veterinary Medicine, 2012, 2, 120-123 /10.4236/ojvm.2012.23020 Published Online September 2012 (http://www.SciRP.org/journal/ojvm) Binary Ethylenimine Inactivated Japanese Encephalitis Virus Antigen Reveals Hemagglutination Dong-Kun Yang , Ha-Hyun Kim, Jin-Ju Nah, Kyung-Woo Lee, Jae-Young Song * Animal, Plant and Fishery Quarantine Inspection Agency (MIFAFF), Anyang, Korea Email: yangdk@korea.kr * Received June 28, 2012; revised July 23, 2012; accepted July 31, 2012 ABSTRACT Severe climate change and global warming may impact significantly on vector-borne disease including Japanese en- cephalitis (JE) infection in human and animals. Thus, veterinary authority requires large quantity of diagnostic tools to survey vector-borne diseases. New producing method having a relation with JE antigen is needed to substitute conven- tional sucrose-acetone extraction method using suckling mouse. So, we developed new manufacturing method using polyethylene glycol (PEG) precipitation. Japanese encephalitis virus (JEV) was propagated in roller bottle containing Vero cell and inactivated with two kinds of inactivating reagents. Viability of the supernatant of bulk containing antigen was checked using Vero cell after inactivation. The supernatant did not show hemagglutination (HA) activity with goose erythrocytes. The antigen inactivated by binary ethylenimine (BEI) and concentrated by PEG precipitation me- thod was found to be 2048 HA, but the antigen inactivated by 0.3% formaldehyde solution and concentrated by PEG precipitation method did not show HA titer. The antigen prepared from mice brain using sucrose-acetone extraction method showed 256 HA titer. This BEI inactivation method does not evoke animal welfare problem and can replace the conventional method that required biological hazardous reagents and suckling mice in preparing HA antigen. This new BEI inactivation method was safe in producing HA an