A nucleocapsid monoclonal antibody based sandwich ELISA for the general detection of both PRRSV-2 and PRRSV-1

Shaohua Sun; Kaili Zhang; Jiajia Zhang; Ping He; Pingping Zhang; Dafu Deng; Chenglin Chi; Sen Jiang; Wanglong Zheng; Nanhua Chen; Jianzhong Zhu

doi:10.1016/j.vetmic.2025.110399

A nucleocapsid monoclonal antibody based sandwich ELISA for the general detection of both PRRSV-2 and PRRSV-1

Vet Microbiol. 2025 Jan 20:302:110399. doi: 10.1016/j.vetmic.2025.110399. Online ahead of print.

Authors

Shaohua Sun¹, Kaili Zhang², Jiajia Zhang³, Ping He⁴, Pingping Zhang⁵, Dafu Deng⁶, Chenglin Chi⁷, Sen Jiang⁸, Wanglong Zheng⁹, Nanhua Chen¹⁰, Jianzhong Zhu¹¹

Affiliations

¹ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: dz120210013@stu.yzu.edu.cn.
² College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: mz120211499@stu.yzu.edu.cn.
³ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: dz120220013@stu.yzu.edu.cn.
⁴ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: mz120221588@stu.yzu.edu.cn.
⁵ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: mz120231738@stu.yzu.edu.cn.
⁶ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: mz120221592@stu.yzu.edu.cn.
⁷ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: dz120230014@stu.yzu.edu.cn.
⁸ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: 008702@yzu.edu.cn.
⁹ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: 007297@yzu.edu.cn.
¹⁰ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: hnchen@yzu.edu.cn.
¹¹ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: jzzhu@yzu.edu.cn.

PMID: 39847872
DOI: 10.1016/j.vetmic.2025.110399

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) causes reproductive failure in sows and respiratory disease in growing pigs, leading to significant economic losses worldwide. Due to the constant mutation and recombination, PRRSV exhibits significant genetic diversity, the general detection of all PRRSV-2 and PRRSV-1 strains is thus needed. In our study, four monoclonal antibodies (mAbs) against PRRSV nucleocapsid (N) protein were generated and the precise and novel B cell epitopes (⁵²KPHF⁵⁵ and ¹⁰⁹HHTVR¹¹³) were identified. The epitope ⁵²KPHF⁵⁵ is highly conserved across all strains of PRRSV-2 lineages and PRRSV-1 subtypes, and the corresponding two mAbs (6D7, 4D12) were selected to develop a sandwich ELISA that was able to detect all tested PRRSV-2 and PRRSV-1 strains. The developed sandwich ELISA demonstrated high specificity, sensitivity and repeatability. In detection of 133 clinical samples, the sandwich ELISA achieved 84.21 % coincidence with the real-time RT-PCR. In conclusion, the mAb based sandwich ELISA can be suitable for detection of potential all PRRSV-2 lineages and PRRSV-1 subtypes, providing a simple, quick and high content method for diagnosis of PRRS.

Keywords: Epitope; Monoclonal antibody; N protein; PRRSV; Sandwich ELISA.