Leonard Limpinsel , Gerhard Dobler , Johannes P. Borde , Julia M. Riehm , Philipp Girl
{"title":"巴伐利亚野猪布鲁氏菌病监测:与世界动物卫生组织列出的血清学检测相比,评估一种新型基于rLPS/ slps的ELISA","authors":"Leonard Limpinsel , Gerhard Dobler , Johannes P. Borde , Julia M. Riehm , Philipp Girl","doi":"10.1016/j.vetmic.2025.110679","DOIUrl":null,"url":null,"abstract":"<div><h3>Introduction</h3><div>Brucellosis is a globally significant zoonotic disease, caused by <em>Brucella spp.</em>, with wildlife reservoirs such as wild boars posing a potential threat to brucellosis-free livestock populations and public health. Despite eradication of brucellosis in domestic animals in Germany, the disease persists in wildlife. Reliable and specific diagnostic tools are essential for effective surveillance.</div></div><div><h3>Material and methods</h3><div>In this study, 149 serum samples from wild boars hunted during the 2023/2024 season in Bavaria (Germany) were analyzed using four serological tests: 1.the rose bengal test (RBT), 2. a conventional sLPS-based ELISA (BMS),3. a novel biwell ELISA (BSI) using both sLPS and rLPS antigens and 4. the complement fixation test (CFT) as gold standard. Sensitivity, specificity, and accuracy of all assays were calculated in comparison to the CFT.</div></div><div><h3>Results</h3><div>Of the 149 samples analyzed, 9 tested positive by CFT, resulting in a seroprevalence of 6.0 % [3.1 %, 11.2 %]in the sampled wild boar population. The BMS-ELISA demonstrated the highest sensitivity (100 %) but moderate specificity (85.0 %), whereas the BSI ELISA showed improved specificity (94.3 %) and accuracy (92.6 %) through combined detection of antibodies against sLPS and rLPS, albeit with lower sensitivity (66.7 %). The RBT performed least favorably with a sensitivity of 55.6 % and specificity of 92.7 %.</div></div><div><h3>Discussion</h3><div>The findings confirm that brucellosis remains endemic among wild boars in Bavaria. While the BSI ELISA shows promise due to its high specificity, its lower sensitivity limits its utility as a stand-alone diagnostic. Cross-reactions in sLPS-based assays highlight the importance of combining antigens for improved test reliability. The varying seroprevalence compared to previous studies underscores the dynamic nature of infection in wildlife populations.</div></div><div><h3>Conclusion</h3><div>A dual-step diagnostic approach – utilizing a sensitive ELISA for screening followed by CFT for confirmation – remains the most effective strategy for wildlife surveillance. The BSI ELISA may serve as a viable alternative in clinical or resource-limited settings. Continued monitoring is crucial to mitigate the zoonotic risk posed by wildlife reservoirs.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"309 ","pages":"Article 110679"},"PeriodicalIF":2.7000,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Brucellosis surveillance in Bavarian wild boar: Evaluation of a novel rLPS/sLPS-based ELISA compared to OIE-listed serological tests\",\"authors\":\"Leonard Limpinsel , Gerhard Dobler , Johannes P. Borde , Julia M. Riehm , Philipp Girl\",\"doi\":\"10.1016/j.vetmic.2025.110679\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Introduction</h3><div>Brucellosis is a globally significant zoonotic disease, caused by <em>Brucella spp.</em>, with wildlife reservoirs such as wild boars posing a potential threat to brucellosis-free livestock populations and public health. Despite eradication of brucellosis in domestic animals in Germany, the disease persists in wildlife. Reliable and specific diagnostic tools are essential for effective surveillance.</div></div><div><h3>Material and methods</h3><div>In this study, 149 serum samples from wild boars hunted during the 2023/2024 season in Bavaria (Germany) were analyzed using four serological tests: 1.the rose bengal test (RBT), 2. a conventional sLPS-based ELISA (BMS),3. a novel biwell ELISA (BSI) using both sLPS and rLPS antigens and 4. the complement fixation test (CFT) as gold standard. Sensitivity, specificity, and accuracy of all assays were calculated in comparison to the CFT.</div></div><div><h3>Results</h3><div>Of the 149 samples analyzed, 9 tested positive by CFT, resulting in a seroprevalence of 6.0 % [3.1 %, 11.2 %]in the sampled wild boar population. The BMS-ELISA demonstrated the highest sensitivity (100 %) but moderate specificity (85.0 %), whereas the BSI ELISA showed improved specificity (94.3 %) and accuracy (92.6 %) through combined detection of antibodies against sLPS and rLPS, albeit with lower sensitivity (66.7 %). The RBT performed least favorably with a sensitivity of 55.6 % and specificity of 92.7 %.</div></div><div><h3>Discussion</h3><div>The findings confirm that brucellosis remains endemic among wild boars in Bavaria. While the BSI ELISA shows promise due to its high specificity, its lower sensitivity limits its utility as a stand-alone diagnostic. Cross-reactions in sLPS-based assays highlight the importance of combining antigens for improved test reliability. The varying seroprevalence compared to previous studies underscores the dynamic nature of infection in wildlife populations.</div></div><div><h3>Conclusion</h3><div>A dual-step diagnostic approach – utilizing a sensitive ELISA for screening followed by CFT for confirmation – remains the most effective strategy for wildlife surveillance. The BSI ELISA may serve as a viable alternative in clinical or resource-limited settings. Continued monitoring is crucial to mitigate the zoonotic risk posed by wildlife reservoirs.</div></div>\",\"PeriodicalId\":23551,\"journal\":{\"name\":\"Veterinary microbiology\",\"volume\":\"309 \",\"pages\":\"Article 110679\"},\"PeriodicalIF\":2.7000,\"publicationDate\":\"2025-08-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Veterinary microbiology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0378113525003141\",\"RegionNum\":2,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Veterinary microbiology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0378113525003141","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Brucellosis surveillance in Bavarian wild boar: Evaluation of a novel rLPS/sLPS-based ELISA compared to OIE-listed serological tests
Introduction
Brucellosis is a globally significant zoonotic disease, caused by Brucella spp., with wildlife reservoirs such as wild boars posing a potential threat to brucellosis-free livestock populations and public health. Despite eradication of brucellosis in domestic animals in Germany, the disease persists in wildlife. Reliable and specific diagnostic tools are essential for effective surveillance.
Material and methods
In this study, 149 serum samples from wild boars hunted during the 2023/2024 season in Bavaria (Germany) were analyzed using four serological tests: 1.the rose bengal test (RBT), 2. a conventional sLPS-based ELISA (BMS),3. a novel biwell ELISA (BSI) using both sLPS and rLPS antigens and 4. the complement fixation test (CFT) as gold standard. Sensitivity, specificity, and accuracy of all assays were calculated in comparison to the CFT.
Results
Of the 149 samples analyzed, 9 tested positive by CFT, resulting in a seroprevalence of 6.0 % [3.1 %, 11.2 %]in the sampled wild boar population. The BMS-ELISA demonstrated the highest sensitivity (100 %) but moderate specificity (85.0 %), whereas the BSI ELISA showed improved specificity (94.3 %) and accuracy (92.6 %) through combined detection of antibodies against sLPS and rLPS, albeit with lower sensitivity (66.7 %). The RBT performed least favorably with a sensitivity of 55.6 % and specificity of 92.7 %.
Discussion
The findings confirm that brucellosis remains endemic among wild boars in Bavaria. While the BSI ELISA shows promise due to its high specificity, its lower sensitivity limits its utility as a stand-alone diagnostic. Cross-reactions in sLPS-based assays highlight the importance of combining antigens for improved test reliability. The varying seroprevalence compared to previous studies underscores the dynamic nature of infection in wildlife populations.
Conclusion
A dual-step diagnostic approach – utilizing a sensitive ELISA for screening followed by CFT for confirmation – remains the most effective strategy for wildlife surveillance. The BSI ELISA may serve as a viable alternative in clinical or resource-limited settings. Continued monitoring is crucial to mitigate the zoonotic risk posed by wildlife reservoirs.
期刊介绍:
Veterinary Microbiology is concerned with microbial (bacterial, fungal, viral) diseases of domesticated vertebrate animals (livestock, companion animals, fur-bearing animals, game, poultry, fish) that supply food, other useful products or companionship. In addition, Microbial diseases of wild animals living in captivity, or as members of the feral fauna will also be considered if the infections are of interest because of their interrelation with humans (zoonoses) and/or domestic animals. Studies of antimicrobial resistance are also included, provided that the results represent a substantial advance in knowledge. Authors are strongly encouraged to read - prior to submission - the Editorials (''Scope or cope'' and ''Scope or cope II'') published previously in the journal. The Editors reserve the right to suggest submission to another journal for those papers which they feel would be more appropriate for consideration by that journal.
Original research papers of high quality and novelty on aspects of control, host response, molecular biology, pathogenesis, prevention, and treatment of microbial diseases of animals are published. Papers dealing primarily with immunology, epidemiology, molecular biology and antiviral or microbial agents will only be considered if they demonstrate a clear impact on a disease. Papers focusing solely on diagnostic techniques (such as another PCR protocol or ELISA) will not be published - focus should be on a microorganism and not on a particular technique. Papers only reporting microbial sequences, transcriptomics data, or proteomics data will not be considered unless the results represent a substantial advance in knowledge.
Drug trial papers will be considered if they have general application or significance. Papers on the identification of microorganisms will also be considered, but detailed taxonomic studies do not fall within the scope of the journal. Case reports will not be published, unless they have general application or contain novel aspects. Papers of geographically limited interest, which repeat what had been established elsewhere will not be considered. The readership of the journal is global.