Development and evaluation of the recombinant BP26 protein-based C-ELISA for human brucellosis diagnosis
IntroductionTimely and accurate diagnosis is crucial for the effective treatment and prevention of brucellosis. Current serological diagnostics, primarily based on lipopolysaccharide (LPS), suffer from cross-reactivity with other Gram-negative bacteria, which limits their specificity. Periplasmic pr...
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| Format: | Article |
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1516915/full |
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| author | Yujia Xie Shaoqing Lin Liping Guo Xinru Qi Shiqi Zhao Qichuan Pei Yixiao Chen Qi Wu Yun Wang Meixue Yao Dehui Yin Dehui Yin |
| author_facet | Yujia Xie Shaoqing Lin Liping Guo Xinru Qi Shiqi Zhao Qichuan Pei Yixiao Chen Qi Wu Yun Wang Meixue Yao Dehui Yin Dehui Yin |
| author_sort | Yujia Xie |
| collection | DOAJ |
| description | IntroductionTimely and accurate diagnosis is crucial for the effective treatment and prevention of brucellosis. Current serological diagnostics, primarily based on lipopolysaccharide (LPS), suffer from cross-reactivity with other Gram-negative bacteria, which limits their specificity. Periplasmic protein 26 (BP26), a highly immunogenic antigen found in Brucella, has emerged as a promising alternative for enhancing diagnostic specificity. This study aimed to develop and evaluate a competitive enzyme-linked immunosorbent assay (C-ELISA) utilizing monoclonal antibodies against BP26 for the diagnosis of human brucellosis, thereby providing a more accurate and specific diagnostic approach.MethodsThe study produced monoclonal antibody (mAb) against the BP26 protein through traditional mouse hybridoma technology and developed the C-ELISA method, and compared with a C-ELISA method based on LPS mAb. The detection performance was validated through the analysis of 190 human serum samples, which included 95 brucellosis serum samples and 95 negative serum samples collected by the Xuzhou Center for Disease Control and Prevention, and a comparative analysis was conducted on the diagnostic efficacy of indirect ELISA for brucellosis using both BP26 and LPS-based methods.ResultsThe BP26 mAb based C-ELISA achieved 100% sensitivity and specificity in detecting human brucellosis, significantly outperforming the C-ELISA based LPS mAb. Furthermore, the accuracy of the indirect enzyme-linked immunosorbent assay (I-ELISA) using BP26 protein was 98.95%, compared to an accuracy of LPS diagnosis was 99.47%. These results indicated that the BP26 mAb can effectively and accurately detected human brucellosis infections.ConclusionThis study successfully developed and evaluated a BP26 protein-based C-ELISA method for diagnosing human brucellosis, establishing a foundation for identifying alternative diagnostic antigens for brucellosis. |
| format | Article |
| id | doaj-art-9fe688b6fc8947f09142e9be862bc5bb |
| institution | Kabale University |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj-art-9fe688b6fc8947f09142e9be862bc5bb2025-01-03T06:46:57ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-01-011510.3389/fmicb.2024.15169151516915Development and evaluation of the recombinant BP26 protein-based C-ELISA for human brucellosis diagnosisYujia Xie0Shaoqing Lin1Liping Guo2Xinru Qi3Shiqi Zhao4Qichuan Pei5Yixiao Chen6Qi Wu7Yun Wang8Meixue Yao9Dehui Yin10Dehui Yin11Jiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, ChinaMedical Research Center and Clinical Laboratory, Zhuhai People’s Hospital (The Affiliated Hospital of Beijing Institute of Technology, Zhuhai Clinical Medical College of Jinan University), Zhuhai, ChinaJiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, ChinaJiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, ChinaJiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, ChinaJiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, ChinaJiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, ChinaJiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, ChinaDepartment of Dermatology, the Affiliated Huai’an Hospital of Xuzhou Medical University, the Second People’s Hospital of Huai’an, Huai’an, ChinaJiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, ChinaJiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, ChinaKey Laboratory of Human Genetics and Environmental Medicine, Xuzhou Medical University, Xuzhou, ChinaIntroductionTimely and accurate diagnosis is crucial for the effective treatment and prevention of brucellosis. Current serological diagnostics, primarily based on lipopolysaccharide (LPS), suffer from cross-reactivity with other Gram-negative bacteria, which limits their specificity. Periplasmic protein 26 (BP26), a highly immunogenic antigen found in Brucella, has emerged as a promising alternative for enhancing diagnostic specificity. This study aimed to develop and evaluate a competitive enzyme-linked immunosorbent assay (C-ELISA) utilizing monoclonal antibodies against BP26 for the diagnosis of human brucellosis, thereby providing a more accurate and specific diagnostic approach.MethodsThe study produced monoclonal antibody (mAb) against the BP26 protein through traditional mouse hybridoma technology and developed the C-ELISA method, and compared with a C-ELISA method based on LPS mAb. The detection performance was validated through the analysis of 190 human serum samples, which included 95 brucellosis serum samples and 95 negative serum samples collected by the Xuzhou Center for Disease Control and Prevention, and a comparative analysis was conducted on the diagnostic efficacy of indirect ELISA for brucellosis using both BP26 and LPS-based methods.ResultsThe BP26 mAb based C-ELISA achieved 100% sensitivity and specificity in detecting human brucellosis, significantly outperforming the C-ELISA based LPS mAb. Furthermore, the accuracy of the indirect enzyme-linked immunosorbent assay (I-ELISA) using BP26 protein was 98.95%, compared to an accuracy of LPS diagnosis was 99.47%. These results indicated that the BP26 mAb can effectively and accurately detected human brucellosis infections.ConclusionThis study successfully developed and evaluated a BP26 protein-based C-ELISA method for diagnosing human brucellosis, establishing a foundation for identifying alternative diagnostic antigens for brucellosis.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1516915/fullBP26 proteinmonoclonal antibodybrucellosiscompetitive enzyme-linked immunosorbent assaydiagnosis |
| spellingShingle | Yujia Xie Shaoqing Lin Liping Guo Xinru Qi Shiqi Zhao Qichuan Pei Yixiao Chen Qi Wu Yun Wang Meixue Yao Dehui Yin Dehui Yin Development and evaluation of the recombinant BP26 protein-based C-ELISA for human brucellosis diagnosis Frontiers in Microbiology BP26 protein monoclonal antibody brucellosis competitive enzyme-linked immunosorbent assay diagnosis |
| title | Development and evaluation of the recombinant BP26 protein-based C-ELISA for human brucellosis diagnosis |
| title_full | Development and evaluation of the recombinant BP26 protein-based C-ELISA for human brucellosis diagnosis |
| title_fullStr | Development and evaluation of the recombinant BP26 protein-based C-ELISA for human brucellosis diagnosis |
| title_full_unstemmed | Development and evaluation of the recombinant BP26 protein-based C-ELISA for human brucellosis diagnosis |
| title_short | Development and evaluation of the recombinant BP26 protein-based C-ELISA for human brucellosis diagnosis |
| title_sort | development and evaluation of the recombinant bp26 protein based c elisa for human brucellosis diagnosis |
| topic | BP26 protein monoclonal antibody brucellosis competitive enzyme-linked immunosorbent assay diagnosis |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1516915/full |
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