Development and optimization of a double antigen sandwich ELISA detecting for Senecavirus A antibodies based on VP2 protein

Jie Chen; Zhengwang Shi; Juncong Luo; Caixia Jia; Xiaoyang Zhang; Juanjuan Wei; Shuaipeng Li; Yuqian Zhu; Tao Xi; Jing Zhou; Yindi He; Xintai Shi; Huanchen Liao; Hong Tian; Haixue Zheng

doi:10.1128/spectrum.02043-24

Microbiology Spectrum (Dec 2024)

Development and optimization of a double antigen sandwich ELISA detecting for Senecavirus A antibodies based on VP2 protein

Jie Chen,
Zhengwang Shi,
Juncong Luo,
Caixia Jia,
Xiaoyang Zhang,
Juanjuan Wei,
Shuaipeng Li,
Yuqian Zhu,
Tao Xi,
Jing Zhou,
Yindi He,
Xintai Shi,
Huanchen Liao,
Hong Tian,
Haixue Zheng

Affiliations

Jie Chen: 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
Zhengwang Shi: 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
Juncong Luo: 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
Caixia Jia: 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
Xiaoyang Zhang: 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
Juanjuan Wei: 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
Shuaipeng Li: 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
Yuqian Zhu: 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
Tao Xi: 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
Jing Zhou: 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
Yindi He: 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
Xintai Shi: 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
Huanchen Liao: 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
Hong Tian: 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
Haixue Zheng: 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

DOI: https://doi.org/10.1128/spectrum.02043-24
Journal volume & issue: Vol. 12, no. 12

Abstract

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ABSTRACT Senecavirus A (SVA) is an emerging viral pathogen that threatens the global swine industry significantly. The major clinical symptoms of SVA-infected animals are vesicular lesions, but various diseases can cause the same symptoms, which makes it difficult to distinguish SVA from other vesicular diseases clinically. The absence of an effective and safe vaccine necessitates the development of a simple, specific, and sensitive serological detection method for SVA antibodies. The VP2 protein of SVA, characterized by high immunogenicity and sequence conservatism, is an essential target for serological diagnosis. In this study, a double-antigen sandwich enzyme-linked immunosorbent assay [ELISA (DAgS-ELISA)] based on VP2 protein expressed by Escherichia coli was established for SVA antibody detection. With a cut-off value of 0.237, this assay demonstrated outstanding performance, showing high sensitivity and sharp specificity, which is manifested in the absence of cross-reaction with classical swine fever virus (CSFV), African swine fever virus (ASFV), pseudorabies virus (PRV), and porcine reproductive and respiratory syndrome virus (PRRSV), and foot-and-mouth disease virus (FMDV) serotype A and O. Additionally, the repeatability of the method is remarkable, as shown by the coefficients variation (CV) of both the intra- and inter-assay below 10%. By detecting 166 clinical sera, it was found that the kappa value of the DAgS-ELISA was 0.78 compared with that of the virus neutralization test (VNT), indicating a high level of consistency. In general, this method has high sensitivity, sharp specificity, remarkable repeatability, sound consistency, and low cost, making it a reliable and effective tool for detecting SVA antibodies.IMPORTANCESVA has rapidly become prevalent in many countries, and its outbreaks have threatened the global swine industry significantly. The major clinical symptoms of SVA-infected animals are vesicular lesions that are similar to other vesicular diseases, making it difficult to distinguish SVA. Currently, no commercial vaccines are available for SVA; therefore, effective diagnosis of SVA infection is vital for its prevention and control. In this study, VP2 protein of SVA was expressed by E. coli, and a double-antigen sandwich enzyme-linked immunosorbent assay [ELISA (DAgS-ELISA)] for SVA antibodies detection was successfully established based on the VP2 protein. The DAgS-ELISA has a high sensitivity, sharp specificity, remarkable repeatability, sound consistency, and low cost for detecting SVA antibodies. Therefore, the DAgS-ELISA established in this study may be a reliable and effective tool for detecting SVA antibodies and may be used to strengthen the monitoring and prevention of SVA epidemic in the long run.

Published in Microbiology Spectrum

ISSN: 2165-0497 (Online)
Publisher: American Society for Microbiology
Country of publisher: United States
LCC subjects: Science: Microbiology
Website: https://journals.asm.org/journal/spectrum

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