Evaluation of twin-arginine translocation substrate proteins as potential antigen candidates for serodiagnosis of brucellosis

IntroductionBrucellosis, 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-argi...

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Main Authors: Yao Wu, Xin Yan, Mingjun Sun, Xiaohan Guo, Jiaqi Li, Xiangxiang Sun, Mengda Liu, Haobo Zhang, Wenlong Nan, Weixing Shao, Fangkun Wang, Xiaoxu Fan, Shufang Sun
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-02-01
Series:Frontiers in Veterinary Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fvets.2025.1398983/full
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author Yao Wu
Yao Wu
Xin Yan
Mingjun Sun
Xiaohan Guo
Jiaqi Li
Xiangxiang Sun
Xiangxiang Sun
Mengda Liu
Mengda Liu
Haobo Zhang
Haobo Zhang
Wenlong Nan
Weixing Shao
Fangkun Wang
Xiaoxu Fan
Xiaoxu Fan
Shufang Sun
author_facet Yao Wu
Yao Wu
Xin Yan
Mingjun Sun
Xiaohan Guo
Jiaqi Li
Xiangxiang Sun
Xiangxiang Sun
Mengda Liu
Mengda Liu
Haobo Zhang
Haobo Zhang
Wenlong Nan
Weixing Shao
Fangkun Wang
Xiaoxu Fan
Xiaoxu Fan
Shufang Sun
author_sort Yao Wu
collection DOAJ
description IntroductionBrucellosis, 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.MethodsIn 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.ResultsFor 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.ConclusionThe findings of this study suggest that Tat substrate proteins serve as promising candidate antigens with significant potential value for the clinical diagnosis of brucellosis.
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spelling doaj-art-64e3002aac3c4eb68e2a99b0e2150af32025-02-05T11:32:17ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692025-02-011210.3389/fvets.2025.13989831398983Evaluation of twin-arginine translocation substrate proteins as potential antigen candidates for serodiagnosis of brucellosisYao Wu0Yao Wu1Xin Yan2Mingjun Sun3Xiaohan Guo4Jiaqi Li5Xiangxiang Sun6Xiangxiang Sun7Mengda Liu8Mengda Liu9Haobo Zhang10Haobo Zhang11Wenlong Nan12Weixing Shao13Fangkun Wang14Xiaoxu Fan15Xiaoxu Fan16Shufang Sun17Laboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaCollege of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaKey Lab of Environment and Health, School of Public Health, Xuzhou Medical University, Xuzhou, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaKey Laboratory of Animal Biosafety Risk Prevention and Control (South), Ministry of Agriculture and Rural Affairs, Qingdao, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaKey Laboratory of Ruminant Infectious Disease Prevention and Control (East), Ministry of Agriculture and Rural Affairs, Qingdao, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaKey Laboratory of Ruminant Infectious Disease Prevention and Control (East), Ministry of Agriculture and Rural Affairs, Qingdao, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaCollege of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaKey Laboratory of Ruminant Infectious Disease Prevention and Control (East), Ministry of Agriculture and Rural Affairs, Qingdao, ChinaLaboratory of Zoonoses, China Animal Health and Epidemiology Center, Qingdao, ChinaIntroductionBrucellosis, 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.MethodsIn 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.ResultsFor 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.ConclusionThe findings of this study suggest that Tat substrate proteins serve as promising candidate antigens with significant potential value for the clinical diagnosis of brucellosis.https://www.frontiersin.org/articles/10.3389/fvets.2025.1398983/fullBrucellatwin-arginine protein translocationtranslocated substratesdiagnosisELISA
spellingShingle Yao Wu
Yao Wu
Xin Yan
Mingjun Sun
Xiaohan Guo
Jiaqi Li
Xiangxiang Sun
Xiangxiang Sun
Mengda Liu
Mengda Liu
Haobo Zhang
Haobo Zhang
Wenlong Nan
Weixing Shao
Fangkun Wang
Xiaoxu Fan
Xiaoxu Fan
Shufang Sun
Evaluation of twin-arginine translocation substrate proteins as potential antigen candidates for serodiagnosis of brucellosis
Frontiers in Veterinary Science
Brucella
twin-arginine protein translocation
translocated substrates
diagnosis
ELISA
title Evaluation of twin-arginine translocation substrate proteins as potential antigen candidates for serodiagnosis of brucellosis
title_full Evaluation of twin-arginine translocation substrate proteins as potential antigen candidates for serodiagnosis of brucellosis
title_fullStr Evaluation of twin-arginine translocation substrate proteins as potential antigen candidates for serodiagnosis of brucellosis
title_full_unstemmed Evaluation of twin-arginine translocation substrate proteins as potential antigen candidates for serodiagnosis of brucellosis
title_short Evaluation of twin-arginine translocation substrate proteins as potential antigen candidates for serodiagnosis of brucellosis
title_sort evaluation of twin arginine translocation substrate proteins as potential antigen candidates for serodiagnosis of brucellosis
topic Brucella
twin-arginine protein translocation
translocated substrates
diagnosis
ELISA
url https://www.frontiersin.org/articles/10.3389/fvets.2025.1398983/full
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