Development and application of double antigen sandwich ELISA to detect the anti-body against N protein of Akabane virus
10.16303/j.cnki.1005-4545.2025.08.04
- VernacularTitle:检测阿卡斑病毒N蛋白抗体的双抗原夹心ELISA方法的建立及应用
- Author:
Hewei CHEN
1
;
Chenyang LU
;
Qing YANG
;
Jiayuan CAO
;
Shaomin QIN
;
Fenglian CHEN
;
Jianmin WU
;
Ying CHEN
;
Ling MA
Author Information
1. 广西壮族自治区兽医研究所广西兽医生物技术重点实验室/农业农村部中国(广西)-东盟跨境动物疫病防控重点实验室,广西南宁 530001;广西大学动物科学技术学院,广西南宁 530004
- Publication Type:Journal Article
- Keywords:
Akabane virus;
AKA;
N protein;
polyclonal antibody;
double antigen sandwich ELISA
- From:
Chinese Journal of Veterinary Science
2025;45(8):1609-1615
- CountryChina
- Language:Chinese
-
Abstract:
To establish a rapid Akabane virus(AKAV)antibody detection method in animals,the re-combination AKAV N protein was expressed by the expression system of Escherichia coli,puri-fied from the supernatant,and used to immunized New Zealand White rabbits.The polyclonal anti-body against AKAV N protein had a titer of 1∶8.192×107 and showed good reactivity.After opti-mizing the reaction conditions,an AKAV double antigen sandwich ELISA antibody detection method was established.This method can detect AKAV antibody in a variety of animals including cattles,sheep and goats.The method had no cross-reactivity with the positive serum of BTV,EHDV,BEV and PRV with the intra-and inter-batch coefficients of variation less than 10％.Com-pared with the AKAV blocking ELISA kit,the sensibility of the method was increased by 8-16 folds with a compliance rate of 93.33％and the κ value of 0.864.These results showed that the double antigen sandwich ELISA displayed strong specificity,high sensitivity and repeatability,which could provide technical suport for detection,the prevention and control of AKAV.
