Establishment and evaluation of a method for detection of ASFV antigen by doub-le-antibody sandwich ELISA
10.16303/j.cnki.1005-4545.2024.08.01
- VernacularTitle:双抗体夹心ELISA检测ASFV抗原方法的建立与评价
- Author:
Qixuan LI
1
;
Huixian YUE
;
Yiqian JIANG
;
Yanyan ZHANG
;
Teng CHEN
;
Shuchao WANG
;
Shoufeng ZHANG
;
Rongliang HU
Author Information
1. 中国农业科学院长春兽医研究所,吉林长春 130122
- Keywords:
African swine fever virus;
hyperimmune serum;
monoclonal antibody;
sandwich ELISA
- From:
Chinese Journal of Veterinary Science
2024;44(8):1579-1584,1592
- CountryChina
- Language:Chinese
-
Abstract:
African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10％.The total co-incidence rate with real-time fluorescence quantitative PCR was 92％(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.
