Development and Evaluation of a DAS-ELISA for Rapid Detection of Tembusu Virus Using Monoclonal Antibodies against the Envelope Protein
(0) Since April 2010,Tembusu virus (TMUV) which is a contagious pathgen of waterfowls,causing symptoms of high fever,loss of appetite and fall in egg production,has been reported in east of China.A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) which detects for Tembusu virus (TMUV) was developed,using two monoclonal antibodies (mAbs) against the TMUV envelope (E) protein.BALB/c mice were immunized with purified recombinant E protein expressed in E.coli.Three hybridoma cell lines designated as 1281,10C6 and 2D2,were screened by cell fusion and indirect ELISA for their ability to recognize different linear epitopes on the E protein,and were characterized subsequently.High-affinity mAbs 1281 and 2D2 were used as capture and detection antibodies,respectively.The reaction conditions for the DAS-ELISA were optimized for TMUV detection.The cross-reactivity of the DAS-ELISA was determined using TMUV,duck plague virus,avian influenza virus subtype H9,Newcastle disease virus,duck hepatitis A virus type l and duck reovirus samples.A total of 191 homogenized tissues of field samples were simultaneously detected by DAS-ELISA and by RT-PCR.The former was found to have a high specificity of 99.1% and a sensitivity of 93.1%.These results reveal a positive coincidence between DAS-ELISA and RT-PCR at a coincidence rate of 95.8%.The method developed in this study can be used for the diagnosis of TMUV infection of duck origin.
TMUV Envelope protein Monoclonal antibody Double Antibody Sandwich ELISA
陈浩 欧全宾 唐熠 高绪慧 刁友祥
College of Animal Science and Technology,Shandong Agricultural University,Tai”an,Shandong 271018,China
国内会议
第五届中国兽药大会(中国畜牧兽医学会动物药品学分会2014年学术年会、中国畜牧兽医学会生物制药学分会暨中国微生物学会兽医微生物学专业委员会联合学术论坛)
沈阳
英文
252-260
2014-09-13（万方平台首次上网日期，不代表论文的发表时间）