Develop an indirect ELISA utilizing gD protein to detect antibodies against bovine herpesvirus type 1

Bovine herpesvirus type 1 (BHV-1) is a highly contagious DNA virus that causes a variety of diseases affecting the reproductive and respiratory tracts. These diseases can reduce the health and production performance of cattle, causing significant economic losses in the cattle industry. The current E...

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Main Authors: Qiang Liu, Ruijin Liang, Xiaoxia Niu, Lingling Jiang, Gang Zhang, Pu Wang, Sinong Zhang, Weifeng Gao, Yujiong Wang, Huichen Guo, Yong Li
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Cellular and Infection Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fcimb.2025.1591304/full
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author Qiang Liu
Ruijin Liang
Xiaoxia Niu
Lingling Jiang
Gang Zhang
Pu Wang
Sinong Zhang
Weifeng Gao
Yujiong Wang
Huichen Guo
Huichen Guo
Yong Li
author_facet Qiang Liu
Ruijin Liang
Xiaoxia Niu
Lingling Jiang
Gang Zhang
Pu Wang
Sinong Zhang
Weifeng Gao
Yujiong Wang
Huichen Guo
Huichen Guo
Yong Li
author_sort Qiang Liu
collection DOAJ
description Bovine herpesvirus type 1 (BHV-1) is a highly contagious DNA virus that causes a variety of diseases affecting the reproductive and respiratory tracts. These diseases can reduce the health and production performance of cattle, causing significant economic losses in the cattle industry. The current ELISA kits used to detect BHV-1 have long lead times and are expensive, and are not suitable for bulk testing on large farms. therefore, there is an urgent need to develop a rapid and cost-effective alternative to the BHV-1 test. In this study, recombinant gD protein was expressed by prokaryotic system, and then used as antigen to immunize New Zealand white rabbits to obtain polyclonal antibodies (pAb). An indirect enzyme-linked immunosorbent assay (iELISA) based on gD protein was established for the detection of BHV-1 antibodies in clinical samples. The optimal cutoff value was determined to be 0.6185 using 60 clinical serum samples. This method had no cross-reaction with other common bovine viruses. The developed iELISA method and commercially available kits were used to detect 60 bovine serum samples, with a concordance rate of 93.3%. In summary, we established a rapid and reliable iELISA method based on gD protein, which is suitable for epidemio-logical monitoring of BHV-1.
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institution OA Journals
issn 2235-2988
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publishDate 2025-05-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Cellular and Infection Microbiology
spelling doaj-art-e7b2849b71f841c6beb03213a278849e2025-08-20T02:16:15ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-05-011510.3389/fcimb.2025.15913041591304Develop an indirect ELISA utilizing gD protein to detect antibodies against bovine herpesvirus type 1Qiang Liu0Ruijin Liang1Xiaoxia Niu2Lingling Jiang3Gang Zhang4Pu Wang5Sinong Zhang6Weifeng Gao7Yujiong Wang8Huichen Guo9Huichen Guo10Yong Li11Key Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaQueen’s University Belfast Joint Institute, China Medical University, Shenyang, ChinaKey Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaKey Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaKey Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaKey Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaKey Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaKey Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaKey Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaKey Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaState Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, ChinaKey Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, ChinaBovine herpesvirus type 1 (BHV-1) is a highly contagious DNA virus that causes a variety of diseases affecting the reproductive and respiratory tracts. These diseases can reduce the health and production performance of cattle, causing significant economic losses in the cattle industry. The current ELISA kits used to detect BHV-1 have long lead times and are expensive, and are not suitable for bulk testing on large farms. therefore, there is an urgent need to develop a rapid and cost-effective alternative to the BHV-1 test. In this study, recombinant gD protein was expressed by prokaryotic system, and then used as antigen to immunize New Zealand white rabbits to obtain polyclonal antibodies (pAb). An indirect enzyme-linked immunosorbent assay (iELISA) based on gD protein was established for the detection of BHV-1 antibodies in clinical samples. The optimal cutoff value was determined to be 0.6185 using 60 clinical serum samples. This method had no cross-reaction with other common bovine viruses. The developed iELISA method and commercially available kits were used to detect 60 bovine serum samples, with a concordance rate of 93.3%. In summary, we established a rapid and reliable iELISA method based on gD protein, which is suitable for epidemio-logical monitoring of BHV-1.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1591304/fullbovine herpesvirus type 1gD proteinprokaryotic expressionindirect ELISAantibody detection
spellingShingle Qiang Liu
Ruijin Liang
Xiaoxia Niu
Lingling Jiang
Gang Zhang
Pu Wang
Sinong Zhang
Weifeng Gao
Yujiong Wang
Huichen Guo
Huichen Guo
Yong Li
Develop an indirect ELISA utilizing gD protein to detect antibodies against bovine herpesvirus type 1
Frontiers in Cellular and Infection Microbiology
bovine herpesvirus type 1
gD protein
prokaryotic expression
indirect ELISA
antibody detection
title Develop an indirect ELISA utilizing gD protein to detect antibodies against bovine herpesvirus type 1
title_full Develop an indirect ELISA utilizing gD protein to detect antibodies against bovine herpesvirus type 1
title_fullStr Develop an indirect ELISA utilizing gD protein to detect antibodies against bovine herpesvirus type 1
title_full_unstemmed Develop an indirect ELISA utilizing gD protein to detect antibodies against bovine herpesvirus type 1
title_short Develop an indirect ELISA utilizing gD protein to detect antibodies against bovine herpesvirus type 1
title_sort develop an indirect elisa utilizing gd protein to detect antibodies against bovine herpesvirus type 1
topic bovine herpesvirus type 1
gD protein
prokaryotic expression
indirect ELISA
antibody detection
url https://www.frontiersin.org/articles/10.3389/fcimb.2025.1591304/full
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