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ORIGINAL RESEARCH article
Front. Vet. Sci.
Sec. Veterinary Infectious Diseases
Volume 12 - 2025 |
doi: 10.3389/fvets.2025.1398983
Evaluation of twin arginine translocation system substrate proteins as potential antigen candidates for serodiagnosis of brucellosis
Provisionally accepted
Yao Wu 1,2
Xin Yan 1* Mingjun Sun 1*
Xiaohan Guo 3 Jiaqi Li 1* Xiangxiang Sun 1,4* Mengda Liu 1,5* Haobo Zhang 1,5* Wenlong Nan 1* Weixing Shao 1*
Fangkun Wang 6* Xiaoxu Fan 1,5* Shufang Sun 1*- 1 Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
- 2 Shandong Agricultural University, Taian, China
- 3 School of Public Health, Xuzhou Medical University, Xuzhou, Jiangsu Province, China
- 4 Key Laboratory of Animal Biosafety Risk Prevention and Control (South), Ministry of Agriculture and Rural Affairs, Qingdao, China
- 5 Key Laboratory of Ruminant Infectious Disease Prevention and Control (East), Ministry of Agriculture and Rural Affairs, Qingdao, China
- 6 College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong Province, China
Brucellosis, an infectious zoonotic disease caused by members of the genus Brucella, results in chronic multi-organ injury. Improving the specificity and sensitivity of serological methods for diagnosing brucellosis necessitates the development of novel diagnostic antigens. The twin-arginine translocation (Tat) pathway is responsible for transporting folded proteins across the cytoplasmic membrane and has been implicated in the virulence of Brucella. Three Tat substrate proteins—L,D-transpeptidase ErfK (A0577), linear amide C-N hydrolase YxeI (A1479), and thioesterase domain-containing protein EntF (B0249) contribute significantly to Brucella virulence. However, the roles of these Tat substrate proteins in diagnosing brucellosis remain unclear. In this study, ErfK, YxeI, and EntF were expressed in prokaryotic cells and utilized as diagnostic antigens. The clinical sera from bovines and sheep diagnosed with brucellosis were analyzed using indirect ELISA with these proteins. For bovine serum, the combined protein group (ErfK + YxeI + EntF) and YxeI demonstrated the highest diagnostic accuracy of 94.23% and 93.58%, respectively. Meanwhile, the combined protein group showed the strongest ability to detect Brucella in sheep serum, achieving an accuracy of 88.10%. Both the combined protein group and YxeI displayed no cross-reactivity with rabbit serum immunized against Yersinia enterocolitica O9, Escherichia coli O157:H7, Mycobacterium tuberculosis, Vibrio cholerae, Legionella, and Salmonella, indicating relatively good specificity. The findings of this study suggest that Tat substrate proteins serve as promising candidate antigens with significant potential value for the clinical diagnosis of brucellosis.
Keywords: Brucella, twin-arginine protein translocation, translocated substrates, diagnosis, ELISA
Received: 11 Mar 2024; Accepted: 20 Jan 2025.
Copyright: © 2025 Wu, Yan, Sun, Guo, Li, Sun, Liu, Zhang, Nan, Shao, Wang, Fan and Sun. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Xin Yan, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
Mingjun Sun, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
Jiaqi Li, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
Xiangxiang Sun, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
Mengda Liu, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
Haobo Zhang, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
Wenlong Nan, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
Weixing Shao, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
Fangkun Wang, College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, Shandong Province, China
Xiaoxu Fan, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
Shufang Sun, Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China
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