Development a high-sensitivity sandwich ELISA for determining antigen content of porcine circovirus type 2 vaccines

Lele Xu; Zhihao Chen; Haoyang Gong; Xiuxiu Pei; Yiyao Zhu; Yuchen Lu; Yumiao Wang; Shifa Nan; Yupeng Yin; Qin Zhao; Yunpeng Fan; Yani Sun; Shuqi Xiao

doi:10.1016/j.jviromet.2024.114954

Development a high-sensitivity sandwich ELISA for determining antigen content of porcine circovirus type 2 vaccines

J Virol Methods. 2024 May 17:328:114954. doi: 10.1016/j.jviromet.2024.114954. Online ahead of print.

Authors

Lele Xu¹, Zhihao Chen¹, Haoyang Gong¹, Xiuxiu Pei¹, Yiyao Zhu¹, Yuchen Lu¹, Yumiao Wang¹, Shifa Nan¹, Yupeng Yin², Qin Zhao², Yunpeng Fan³, Yani Sun⁴, Shuqi Xiao⁵

Affiliations

¹ College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China; State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
² College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
³ College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China. Electronic address: ypfan@nwsuaf.edu.cn.
⁴ College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China. Electronic address: sunyani@nwsuaf.edu.cn.
⁵ State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China. Electronic address: shqxiaojd@126.com.

PMID: 38763359
DOI: 10.1016/j.jviromet.2024.114954

Abstract

Porcine circovirus type 2 (PCV2) is intensely prevalent in global pig farms. The PCV2 vaccine is an important means of preventing and controlling PCV2. The quality control of PCV2 vaccines is predominantly based on detection techniques such as animal testing and neutralizing antibody titration. Measuring the content of effective proteins in vaccines to measure vaccine efficacy is an excellent alternative to traditional methods, which can greatly accelerate the development speed and testing time of vaccines. In this study, we screened a monoclonal antibody (mAb) that can effectively recognize not only the exogenous expression of PCV2 Cap protein but also PCV2 virus. The double antibody sandwich ELISA (DAS-ELISA) was developed using this mAb that specifically recognize PCV2 Cap. The minimum protein content detected by this method is 3.5 ng/mL. This method can be used for the quality control of PCV2 inactivated vaccine and subunit vaccine, and the detection results are consistent with the results of mice animal experiments. This method has the advantages of simple operation, good sensitivity, high specificity and wide application. It can detect the effective antigen Cap protein content of various types of PCV2 vaccines, which not only shorten the vaccine inspection time but also save costs.

Keywords: DAS-ELISA; Porcine circovirus type 2; Quality control; Vaccine.