间接酶联免疫吸附法（ELISA）检测羔羊和母羊血清中乳糖不良链球菌亚种抗体的建立、验证和应用

IF 2.2 3区农林科学 Q1 VETERINARY SCIENCES

Research in veterinary science Pub Date : 2025-03-03 DOI:10.1016/j.rvsc.2025.105604

L.P. Jackson , D. Timofte , K.T. Ballingall , J.S. Duncan

{"title":"间接酶联免疫吸附法（ELISA）检测羔羊和母羊血清中乳糖不良链球菌亚种抗体的建立、验证和应用","authors":"L.P. Jackson , D. Timofte , K.T. Ballingall , J.S. Duncan","doi":"10.1016/j.rvsc.2025.105604","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><div>Neonatal infectious arthritis (NIA) is a bacterial disease of lambs caused by <em>Streptococcus dysgalactiae sub species dysgalactiae</em> (SDSD). NIA negatively impacts animal welfare; whilst its control entails widespread antibiotic use. The aims of this study are to develop, validate and apply an indirect, ELISA assay to inform SDSD vaccine development and epidemiology studies.</div></div><div><h3>Methods</h3><div>The antigen (inactivated isolate of SDSD) was incubated with sera from SDSD infected sheep, negative control sheep and serum from sheep flocks with outbreaks of SDSD NIA. SDSD antibody was detected with rabbit anti-sheep IgG conjugated with horseradish peroxidase. Antibody levels were estimated using peroxidase substrate and detection of colour change.</div><div>Test performance was determined by receiver operating characteristic (ROC) curve analysis and by comparison with the animals' concurrent infection status, as determined by a Real Time PCR assay for SDSD bacterial DNA.</div></div><div><h3>Results</h3><div>Using ROC curve analysis, a median test sensitivity of 85.71 % (95 % CI, 77.51–93.91) and a median test specificity of 73.68 % (95 % CI, 53.89–93.48), with a positive predictive value of 92.31 % (95 % CI, 86.07–98.55), and a negative predictive value of 58.33 % (95 % CI, 36.16–80.50).</div><div>ELISA test performance was reduced when serum ELISA status (<em>n</em> = 362 sheep) was compared with concurrent SDSD infection status. The SDSD ELISA sensitivity was 39.74 % (95 % CI, 28.83–51.46), the specificity 60.92 % (95 % CI, 54.97–66.63), the PPV 21.83 % (95 % CI, 15.34–29.53), and the NPV was 78.64 % (95 % CI, 72.62–83.86).</div></div><div><h3>Conclusion</h3><div>This ELISA assay could be applied to estimate flock sero-prevalences for SDSD and changing SDSD antibody titres in vaccine trials.</div></div>","PeriodicalId":21083,"journal":{"name":"Research in veterinary science","volume":"187 ","pages":"Article 105604"},"PeriodicalIF":2.2000,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"The development, validation and application of an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Streptococcus dysgalactiae subspecies dysgalactiae in lamb and ewe sera\",\"authors\":\"L.P. Jackson , D. Timofte , K.T. Ballingall , J.S. Duncan\",\"doi\":\"10.1016/j.rvsc.2025.105604\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><div>Neonatal infectious arthritis (NIA) is a bacterial disease of lambs caused by <em>Streptococcus dysgalactiae sub species dysgalactiae</em> (SDSD). NIA negatively impacts animal welfare; whilst its control entails widespread antibiotic use. The aims of this study are to develop, validate and apply an indirect, ELISA assay to inform SDSD vaccine development and epidemiology studies.</div></div><div><h3>Methods</h3><div>The antigen (inactivated isolate of SDSD) was incubated with sera from SDSD infected sheep, negative control sheep and serum from sheep flocks with outbreaks of SDSD NIA. SDSD antibody was detected with rabbit anti-sheep IgG conjugated with horseradish peroxidase. Antibody levels were estimated using peroxidase substrate and detection of colour change.</div><div>Test performance was determined by receiver operating characteristic (ROC) curve analysis and by comparison with the animals' concurrent infection status, as determined by a Real Time PCR assay for SDSD bacterial DNA.</div></div><div><h3>Results</h3><div>Using ROC curve analysis, a median test sensitivity of 85.71 % (95 % CI, 77.51–93.91) and a median test specificity of 73.68 % (95 % CI, 53.89–93.48), with a positive predictive value of 92.31 % (95 % CI, 86.07–98.55), and a negative predictive value of 58.33 % (95 % CI, 36.16–80.50).</div><div>ELISA test performance was reduced when serum ELISA status (<em>n</em> = 362 sheep) was compared with concurrent SDSD infection status. The SDSD ELISA sensitivity was 39.74 % (95 % CI, 28.83–51.46), the specificity 60.92 % (95 % CI, 54.97–66.63), the PPV 21.83 % (95 % CI, 15.34–29.53), and the NPV was 78.64 % (95 % CI, 72.62–83.86).</div></div><div><h3>Conclusion</h3><div>This ELISA assay could be applied to estimate flock sero-prevalences for SDSD and changing SDSD antibody titres in vaccine trials.</div></div>\",\"PeriodicalId\":21083,\"journal\":{\"name\":\"Research in veterinary science\",\"volume\":\"187 \",\"pages\":\"Article 105604\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2025-03-03\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Research in veterinary science\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0034528825000785\",\"RegionNum\":3,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"VETERINARY SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Research in veterinary science","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0034528825000785","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"VETERINARY SCIENCES","Score":null,"Total":0}

引用次数: 0

摘要

新生儿感染性关节炎（NIA）是由乳糖分泌不良链球菌亚种（SDSD）引起的羔羊细菌性疾病。NIA对动物福利产生负面影响；而控制它需要广泛使用抗生素。本研究的目的是开发、验证和应用一种间接的ELISA检测方法，为SDSD疫苗的开发和流行病学研究提供信息。方法将抗原（SDSD灭活分离物）与SDSD感染羊血清、阴性对照羊血清和SDSD NIA暴发羊血清孵育。用辣根过氧化物酶偶联兔抗羊IgG检测SDSD抗体。使用过氧化物酶底物和检测颜色变化来估计抗体水平。通过受试者工作特征（ROC）曲线分析和与动物并发感染状态的比较来确定测试性能，通过实时PCR检测SDSD细菌DNA来确定。结果采用ROC曲线分析，中位检测灵敏度为85.71% (95% CI, 77.51 ~ 93.91)，中位检测特异性为73.68% (95% CI, 53.89 ~ 93.48)，阳性预测值为92.31% (95% CI, 86.07 ~ 98.55)，阴性预测值为58.33% （95% CI, 36.16 ~ 80.50）。将血清ELISA状态（n = 362只羊）与并发SDSD感染状态进行比较，降低了ELISA检测性能。SDSD ELISA的敏感性为39.74% (95% CI, 28.83 ~ 51.46)，特异性为60.92% (95% CI, 54.97 ~ 66.63)， PPV为21.83% (95% CI, 15.34 ~ 29.53)， NPV为78.64% （95% CI, 72.62 ~ 83.86）。结论该方法可用于疫苗试验中鸡群血清SDSD患病率及SDSD抗体效价的变化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

The development, validation and application of an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Streptococcus dysgalactiae subspecies dysgalactiae in lamb and ewe sera

Background

Neonatal infectious arthritis (NIA) is a bacterial disease of lambs caused by Streptococcus dysgalactiae sub species dysgalactiae (SDSD). NIA negatively impacts animal welfare; whilst its control entails widespread antibiotic use. The aims of this study are to develop, validate and apply an indirect, ELISA assay to inform SDSD vaccine development and epidemiology studies.

Methods

The antigen (inactivated isolate of SDSD) was incubated with sera from SDSD infected sheep, negative control sheep and serum from sheep flocks with outbreaks of SDSD NIA. SDSD antibody was detected with rabbit anti-sheep IgG conjugated with horseradish peroxidase. Antibody levels were estimated using peroxidase substrate and detection of colour change.

Test performance was determined by receiver operating characteristic (ROC) curve analysis and by comparison with the animals' concurrent infection status, as determined by a Real Time PCR assay for SDSD bacterial DNA.

Results

Using ROC curve analysis, a median test sensitivity of 85.71 % (95 % CI, 77.51–93.91) and a median test specificity of 73.68 % (95 % CI, 53.89–93.48), with a positive predictive value of 92.31 % (95 % CI, 86.07–98.55), and a negative predictive value of 58.33 % (95 % CI, 36.16–80.50).

ELISA test performance was reduced when serum ELISA status (n = 362 sheep) was compared with concurrent SDSD infection status. The SDSD ELISA sensitivity was 39.74 % (95 % CI, 28.83–51.46), the specificity 60.92 % (95 % CI, 54.97–66.63), the PPV 21.83 % (95 % CI, 15.34–29.53), and the NPV was 78.64 % (95 % CI, 72.62–83.86).

Conclusion

This ELISA assay could be applied to estimate flock sero-prevalences for SDSD and changing SDSD antibody titres in vaccine trials.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Research in veterinary science 农林科学-兽医学

CiteScore

4.40

自引率

4.20%

发文量

312

审稿时长

75 days

期刊介绍： Research in Veterinary Science is an International multi-disciplinary journal publishing original articles, reviews and short communications of a high scientific and ethical standard in all aspects of veterinary and biomedical research. The primary aim of the journal is to inform veterinary and biomedical scientists of significant advances in veterinary and related research through prompt publication and dissemination. Secondly, the journal aims to provide a general multi-disciplinary forum for discussion and debate of news and issues concerning veterinary science. Thirdly, to promote the dissemination of knowledge to a broader range of professions, globally. High quality papers on all species of animals are considered, particularly those considered to be of high scientific importance and originality, and with interdisciplinary interest. The journal encourages papers providing results that have clear implications for understanding disease pathogenesis and for the development of control measures or treatments, as well as those dealing with a comparative biomedical approach, which represents a substantial improvement to animal and human health. Studies without a robust scientific hypothesis or that are preliminary, or of weak originality, as well as negative results, are not appropriate for the journal. Furthermore, observational approaches, case studies or field reports lacking an advancement in general knowledge do not fall within the scope of the journal.