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抗生物素蛋白-生物素重组核蛋白竞争ELISA法对绵羊和山羊小反刍兽疫血清学监测与监测的比较诊断效能

Comparative diagnostic efficacy of Avidin-Biotin recombinant nucleoprotein competitive ELISA for serosurveillance and monitoring of peste des petits ruminants in sheep and goats.

作者信息

Balamurugan V, Bokade Prajakta P, Kumar K Vinod, SowjanyaKumari S, Nagalingam M, Hemadri D, Shome B R

机构信息

Indian Council of Agricultural Research -National Institute of Veterinary Epidemiology and Disease Informatics (ICAR-NIVEDI), Yelahanka, Bengaluru 560 064, Karnataka, India.

Indian Council of Agricultural Research -National Institute of Veterinary Epidemiology and Disease Informatics (ICAR-NIVEDI), Yelahanka, Bengaluru 560 064, Karnataka, India.

出版信息

J Immunol Methods. 2023 Jan;512:113409. doi: 10.1016/j.jim.2022.113409. Epub 2022 Dec 17.

Abstract

In this study extensive evaluation of Avidin-Biotin recombinant nucleoprotein competitive ELISA (ABrC-ELISA) was carried out by mass screening of a large number of sera to make use of this assay for serosurveillance and seromonitoring of peste des petits ruminants (PPR) in sheep and goats to evaluate its diagnostic efficacy value and strengthen findings associated with the assay. The recombinant PPR virus (PPRV) nucleoprotein was over-expressed in E. coli, Ni-NTA affinity-purified, and characterized and used as coating diagnostic antigen in ABrC-ELISA, and evaluated using the field sera from animals. On evaluation of the diagnostic performance or efficacy of this assay using the pre-vaccinated and post-vaccinated sera of sheep and goats (n = 1437), the ABrC-ELISA showed a relative diagnostic sensitivity of 87.2% (95% CI: 84.1-90%) and diagnostic specificity of 92.0% (95% CI: 90-93.7%), against well-established existing indigenous H protein-specific PPR competitive ELISA kit with an accuracy of 90.1% (95% CI: 88.5-91.7%) and good or substantial agreement of Cohen's Kappa value of 0.79 ± 0.017 SE (95% CI: 0.76 to 0.82). These findings suggest that the ABrC-ELISA is a potential additional diagnostic tool of a rapid, sensitive, and specific assay for the detection of the PPRV nucleoprotein antibodies in sera of sheep and goats. This PPR Ab Chek kit can be used extensively under field conditions for serosurveillance, and seromonitoring of PPR in sheep and goats at the eradication /post-eradication phase in disease-controlled countries or PPR non-enzootic countries.

摘要

在本研究中,通过对大量血清进行大规模筛查,对抗生物素蛋白-生物素重组核蛋白竞争ELISA(ABrC-ELISA)进行了广泛评估,以利用该检测方法对绵羊和山羊的小反刍兽疫(PPR)进行血清学监测和血清学监测,评估其诊断效能值,并强化与该检测方法相关的研究结果。重组PPR病毒(PPRV)核蛋白在大肠杆菌中过表达,经镍-亚氨基二乙酸亲和纯化、表征后,用作ABrC-ELISA中的包被诊断抗原,并使用动物的现场血清进行评估。在使用绵羊和山羊(n = 1437)的免疫前和免疫后血清评估该检测方法的诊断性能或效能时,ABrC-ELISA相对于成熟的现有本土H蛋白特异性PPR竞争ELISA试剂盒,显示出相对诊断敏感性为87.2%(95% CI:84.1-90%),诊断特异性为92.0%(95% CI:90-93.7%),准确性为90.1%(95% CI:88.5-91.7%),Cohen's Kappa值为0.79±0.017 SE(95% CI:0.76至0.82),具有良好或高度一致性。这些研究结果表明,ABrC-ELISA是一种潜在的额外诊断工具,可用于快速、灵敏且特异的检测绵羊和山羊血清中的PPRV核蛋白抗体。这种PPR Ab Chek试剂盒可在现场条件下广泛用于疾病控制国家或PPR非疫区在根除/根除后阶段对绵羊和山羊进行PPR的血清学监测和血清学监测。

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