Prokaryotic expression of HN gene of bovine parainfluenza virus type 3 and the establishment of indirect ELISA method / 病毒学报
Chinese Journal of Virology
;
(6): 23-28, 2012.
Artigo
em Chinês
| WPRIM
| ID: wpr-354776
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 150; the blocking reagent was 5% skimmed milk; the blocking time was 60 min at 37 degrees C; the second antibody concentration was 110 000; 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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Sangue
/
Ensaio de Imunoadsorção Enzimática
/
Proteína HN
/
Western Blotting
/
Vírus da Parainfluenza 3 Bovina
/
Alergia e Imunologia
/
Escherichia coli
/
Genética
/
Métodos
/
Anticorpos Antivirais
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Virology
Ano de publicação:
2012
Tipo de documento:
Artigo
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