논문 상세보기
Improved solid-phase competition enzyme-linked immunosorbent assay for the specific detection of antibodies against porcine parvovirus based on VP2 protein expressed in baculovirus KCI 등재
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/411981
pp.158-165
A solid-phase competition enzyme-linked immunosorbent assay (ELISA), recombinant VP2 (rVP2) protein, and monoclonal antibody (mAb) were developed for the specific and sensitive detection of porcine parvovirus (PPV) antibodies in pig sera. A total of 1,544 sera samples were collected from breeding pig farms located in the Gyeongsangbuk-do Province in the Republic of Korea. The optimal operating conditions of SC-ELISA were as follows. The concentration of rVP2 proteins coated on the wells was 4 μg/mL, the swine sera were diluted 1:2, and the HRP-conjugated PPV VP2 mAb (9A8 clone) was used at 500 ng/mL. These results suggest that the SC-rVP-ELISA assay may be a valuable alternative to the current diagnostic tools used to detect PPV-specific monoclonal antibodies and broadly monitor PPV infections in domestic pigs at different breeding stages.
MATERIALS AND METHODS
Viruses and cells culture
Serum samples in the field
Production and characterization of anti-PPV mAb
Construction of recombinant plasmids and Bacmid
Expression and purification of the recombinant VP2proteins
Western blot analysis
HA and HI tests
VP2 protein assay
Optimization of the SC-ELISA
Comparison of the analytical performance betweenthe SC-ELISA, commercial ELISA, and HI tests
Statistical analyses
RESULTS
DISCUSSION
ACKNOWLEDGEMENTS
