title: Preparation of chicken IgY against recombinant E2 protein of bovine viral diarrhea virus (BVDV) and development of ELISA and ICA for BVDV detection.
abstract: Bovine viral diarrhea virus (BVDV) infects cattle and may lead to persistent infection (PI). The PI animals harbor BVDV throughout their life and become immune tolerant against BVDV. Thus, diagnosis of this virus in herd is highly important. Recombinant E2 protein expression (using pET-32a in Escherichia coli) was confirmed by SDS-PAGE and Western blotting; then purified by Ni+affinity chromatography. Chickens were immunized with BVDV-E2 protein, and IgY antibodies were extracted from egg yolk by PEG-6000. The peak titer of anti-BVDV-E2-IgY was 1:128,000 after the fifth immunization. IgY-based enzyme-linked immuno sorbent assay (ELISA) and immunochromatographic assay (ICA) were further developed. Coincidence of ELISA and ICA test with RT-PCR was 95.45 and 90.91%, respectively. The anti-BVDV-E2 IgY could be used in routine screening of BVDV infection. Besides, it can also be applicable while licensing and/or using live vaccines; screening of imported products containing bovine serum and strong surveillance of BVDV outbreaks.
authors.affiliation: a College of Veterinary Medicine , Northwest A&F University , Yangling , China.|b College of Veterinary Medicine , Xinjiang Agricultural University , Urumqi , China.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Zhang
authors.affiliation: a College of Veterinary Medicine , Northwest A&F University , Yangling , China.
authors.forename: Thirumalai
authors.initials: T
authors.lastname: Diraviyam
authors.affiliation: c Laipson Biological Information Technology Co., LTD , Luoyang , China.
authors.forename: Xiumei
authors.initials: X
authors.lastname: Li
authors.affiliation: b College of Veterinary Medicine , Xinjiang Agricultural University , Urumqi , China.
authors.forename: Gang
authors.initials: G
authors.lastname: Yao
authors.affiliation: d PSG College of Arts & Science , Coimbatore , India.
authors.forename: Antonysamy
authors.initials: A
authors.lastname: Michael
mesh_terms: D000818:Animals; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods*; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007136:Immunoglobulins / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: C016512:IgY; D007136:Immunoglobulins; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C076551:gp53, bovine viral diarrhea virus
keywords: E2 recombinant protein; ELISA; bovine viral diarrhea virus (BVDV); egg yolk antibody (IgY); immunochromatographic assay (ICA)