Development and preliminary application of an indirect ELISA to detect infectious bovine rhinotracheitis virus using recombinant glycoprotein D of IBRV strain SD [Enfeksiyöz bovine rhinotracheitis virusun tespitinde IBRV suş SD rekombinant glikoprotein D kullanılarak İndirekt ELISA yönteminin geliştirilmesi ve uygulanması]

Glycoprotein d (gd) is the major structural protein of infectious bovine rhinotracheitis virus (ibrv). it can induce both humoral and cellular immunity thus it is a preferable protein for ibr diagnostic reagent. regarding to dnastar analysis,the major antigenic region of the gd fragment was amplified by pcr using the ibrv genomic dna as a template,and subsequenly constructed into the recombinant plasmid pet32a-gd. the fusion protein was expressed upon iptg induction. the fusion protein was purified by immobilized ni ion affinity chromatography with a ni-nta kit after verification by sds-page and western-blotting analysis,then was utilized as a coating antigen to detect antibodies to infectious bovine rhinotracheitis virus (ibrv) in an indirect elisa method. cross-reactivity examinations have showed that the recombinant antigen had no cross reaction with positive sera of other common viral diseases (bovine ephemeral fever,bovine viral diarrhea-mucosal disease,calf diarrhea,bovine intestinal virus infection,bovine coronavirus disease) which indicating a strong specificity. application of this diagnostic method in 1315 clinical serum samples displayed an antibody positive rate of 23.7% (311/1315),with respect to a coincidence rate of 96.8% as compared to a commercialized ibrv whole virus elisa. our method is stable and sensitive hence provides a quick and convenient serological diagnosis favoring epidemiology and disease identification of domestic ibr. © 2016,veteriner fakultesi dergisi. all rights reserved.