Efficiency of recombinant Ybgf in a double antigen-ELISA for the detection of Coxiella antibodies in ruminants

IF 1.9 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Veterinary and Animal Science Pub Date : 2024-06-02 DOI:10.1016/j.vas.2024.100366
Gianmarco Ferrara , Barbara Colitti , Flores-Ramires Gabriela , Sergio Rosati , Giuseppe Iovane , Ugo Pagnini , Serena Montagnaro
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Abstract

Q fever is a zoonosis whose main reservoirs are domestic ruminants. Surveillance in these species is carried out mainly with serological tests, which, however, have limited diagnostic performance, and their manufacturing requires laboratories equipped with high biosafety requirements for antigen production. Recombinant ELISAs do not depend on these requirements and, being based on a single antigen, can reduce potential false positivity by identifying antibodies specific to a phase of infection. The aim of this study was to apply a new technology (dual antigen test) to a recombinant protein (Ybgf), an antigen produced in recombinant form and already used in previous studies for the design of an indirect ELISA. The successfully produced recombinant antigen was used to coat 96-well plates and, at the same time, another antigen aliquot was conjugated with HRP to obtain an HRP-conjugated Ybgf. After setting the test conditions, the results obtained with the recombinant double antigen test were compared with those obtained with a commercial assay (considered as reference assay) testing a total of 514 ruminant samples (280 goats and 234 cattle). A concordance of 86.2 and a Cohen's Kappa value of 0.72 were obtained, with no significant difference between the two species tested. Notably, the test proved to be highly specific, having correctly identified 250 out of 253 animals. This research represents an additional effort to use recombinant antigens to enhance serological methods in veterinary medicine. In a “one-health scenario”, improving the performance of serological tests used in veterinary practice also means improving the surveillance of this infection.

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重组 Ybgf 在双抗原-ELISA 中检测反刍动物体内柯西氏杆菌抗体的效率
Q 热是一种人畜共患病,其主要传染源是家养反刍动物。对这些物种的监测主要通过血清学检测进行,但这种检测方法的诊断性能有限,而且其生产需要实验室配备对抗原生产有较高生物安全要求的设备。重组酶联免疫吸附试验不需要满足这些要求,而且基于单一抗原,可以通过识别感染某一阶段的特异性抗体来减少潜在的假阳性。本研究的目的是将新技术(双抗原检测)应用于重组蛋白(Ybgf),这是一种以重组形式生产的抗原,之前的研究已将其用于间接 ELISA 的设计。成功制备的重组抗原被用于涂布 96 孔板,与此同时,另一份抗原等量物与 HRP 相结合,得到 HRP 结合的 Ybgf。设定检测条件后,将重组双抗原检测法与商业检测法(参考检测法)的检测结果进行了比较,共检测了 514 份反刍动物样本(280 份山羊样本和 234 份牛样本)。结果表明,两种检测方法的一致性为 86.2,Cohen's Kappa 值为 0.72,无明显差异。值得注意的是,该测试的特异性很高,在 253 头动物中正确识别了 250 头。这项研究是利用重组抗原改进兽医血清学方法的又一努力。在 "单一健康方案 "中,提高兽医实践中使用的血清学测试的性能也意味着改进对这种感染的监测。
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来源期刊
Veterinary and Animal Science
Veterinary and Animal Science Veterinary-Veterinary (all)
CiteScore
3.50
自引率
0.00%
发文量
43
审稿时长
47 days
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