Development and Preliminary Application of an Indirect ELISA to Detect Infectious Bovine Rhinotracheitis Virus Using Recombinant Glycoprotein D of IBRV Strain SD
文献类型： 外文期刊
作者： Song, Lingling 1 ; Zhang, Hui 2 ; Hou, Peili 2 ; Wang, Hongmei 2 ; Zhao, Guimin 2 ; Xia, Xianzhu 3 ; He, Hongbin 2 ;
作者机构： 1.Shihezi Univ, Coll Anim Sci & Technol, Shihezi 832003, Peoples R China
2.Shandong Acad Agr Sci, Dairy Cattle Res Ctr, Jinan 250100, Peoples R China
3.Acad Mil Med Sci, Inst Mil Vet, Changchun 130122, Peoples R China
关键词： gD;Indirect ELISA;Infectious bovine rhinotracheitis virus;Serology
期刊名称：KAFKAS UNIVERSITESI VETERINER FAKULTESI DERGISI （ 影响因子：0.685； 五年影响因子：0.655 ）
ISSN： 1300-6045
年卷期： 2016 年 22 卷 4 期
页码：
收录情况： SCI
摘要： 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.
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