title: [Prokaryotic expression of HN gene of bovine parainfluenza virus type 3 and the establishment of indirect ELISA method].
abstract: The prokaryotic expression plasmid pQE30-HN of hemagglutinin-neuraminidase (HN) protein gene of bovine parainfluenza virus type 3 (BPIV3) strain HJ-1 was expressed by IPTG induction in E. coli XL1Blue. The recombinant HN protein(rHN) was purified by electroeluting method, and used as coated antigen. An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect the antibody valence of BPIV3. The best working conditions of ELISA were as follows: the antigen concentration was 6 microg/mL; the serum dilution was 1:50; the blocking reagent was 5% skimmed milk; the blocking time was 60 min at 37 degrees C; the second antibody concentration was 1:10 000; The cut-off value was 0.30. The method revealed a good specificity, no cross-reaction to the positive sera of BCV, IBRV or BRSV was observed. We applied the method to detect 323 serum samples of dairy cow in Heilongjiang Province, the seropositivity rate of BPIV3 was about 58%. The indirect ELISA established provided a technological basis for the development of ELISA kit.
authors.affiliation: Animal Technology College, Heilongjiang Bayi Agricultural University, Daqing 163319, China.
authors.forename: Yu-Long
authors.initials: YL
authors.lastname: Zhou
authors.forename: Ya-Chao
authors.initials: YC
authors.lastname: Ren
authors.forename: Zhan-Bo
authors.initials: ZB
authors.lastname: Zhu
authors.forename: Xi-Lin
authors.initials: XL
authors.lastname: Hou
authors.forename: Mi
authors.initials: M
authors.lastname: Wang
authors.forename: Jing
authors.initials: J
authors.lastname: Geng
authors.forename: Fan-Ze
authors.initials: FZ
authors.lastname: Piao
authors.forename: Sen
authors.initials: S
authors.lastname: Li
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D015153:Blotting, Western; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015707:HN Protein / Q000235:genetics*; D029101:Parainfluenza Virus 3, Bovine / Q000235:genetics* / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D015707:HN Protein