拉霍氏鸟疫暴露间接ELISA检测方法的建立与现场验证

IF 1.4 Q3 PARASITOLOGY

Veterinary parasitology, regional studies and reports Pub Date : 2025-08-09 DOI:10.1016/j.vprsr.2025.101327

Xiaolin Xu , Xiangfen Yuan , Huiyu Wang , Ailixire Maimaiti , Yufang Kong , Jizhou Lv , Shaoqiang Wu

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引用次数: 0

摘要

背景：拉霍氏蜱是许多蜱传疾病的主要媒介。有效的蜱监测有助于预防蜱传疾病。蜱虫叮咬的直接监测方法是不切实际的，开发一种间接方法来检测动物样本中针对蜱虫唾液蛋白的特异性抗体将提供一种更方便的监测和诊断工具。以前的研究已经证实，一些唾液腺蛋白可以作为蜱叮咬的检测标记。方法表达并纯化lahorensis重组BTSP蛋白（rOl-BTSP）。对roll - btsp进行了序列比对分析，并通过Western blot分析验证了其作为lahorensi暴露的血清学检测靶点。在此基础上，通过优化抗原包被浓度、血清稀释度等关键参数，建立了一种检测绵羊拉霍弧菌叮咬的ELISA检测方法（S-iELISA）。评价S-iELISA法的敏感性和特异性。并进一步应用S-iELISA对新疆和内蒙古地区455份绵羊血清进行了检测。结果序列同源性分析显示，Ol-BTSP与硬蜱和软蜱同源物的氨基酸同源性仅为30.5%。结果表明，Ol-BTSP对咬伤小鼠血清具有特异性免疫反应性，而跨种抗体检测无反应性，证实了Ol-BTSP对拉霍弧菌暴露的诊断潜力。开发的S-iELISA具有出色的诊断准确性，在验证研究中灵敏度为93.3% (95% CI: 70.2 - 99.7%)，特异性为96.7% （95% CI: 82.8 - 99.9%）。新疆和内蒙古地区血清暴露拉霍氏弧菌的血清阳性率分别为10.45%和3.09%。结论本研究建立的S-iELISA方法是一种可靠的检测和监测绵羊lahorensis暴露的工具。基于rOl-BTSP的血清学诊断方法对加强拉霍朗氏虫传播疾病的监测和管理具有重要意义。

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Development and field validation of an indirect ELISA for serosurveillance of Ornithodoros lahorensis exposure

Background

Ornithodoros lahorensis is the primary vector of numerous tick-borne diseases. Effective tick surveillance could facilitate the prevention of tick-borne diseases. Direct surveillance methods for tick bites are impractical, the development of an indirect approach to detect specific antibodies against tick salivary proteins in animal samples would provide a more convenient surveillance and diagnostic tool. Previous studies have confirmed that some salivary gland proteins can be used as detection markers for tick bites.

Methods

The recombinant BTSP protein of O. lahorensis (rOl-BTSP) was expressed and purified. rOl-BTSP was subjected to sequence alignment analysis, and validated as a serological detection target for O. lahorensi exposure by Western blot analysis. Then, a novel ELISA method for detecting O. lahorensis bites in sheep (S-iELISA) was developed by optimizing critical parameters, including antigen coating concentration, serum dilution. The sensitivity and specificity of the S-iELISA method were evaluated. And the S-iELISA were further applied to test 455 sheep serum samples from Xinjiang and Inner Mongolia in China.

Results

Sequence homology analysis revealed that Ol-BTSP exhibited only 30.5 % amino acid identity with its homologs from both hard ticks and soft ticks. And the serum from mice bitten by O. lahorensis exhibited specific immunoreactivity with rOl-BTSP, while the cross-species antibody tests showed no reactivity, confirming the diagnostic potential of Ol-BTSP for detecting O. lahorensis exposure. The developed S-iELISA exhibited excellent diagnostic accuracy, with a sensitivity of 93.3 % (95 % CI: 70.2–99.7 %) and a specificity of 96.7 % (95 % CI: 82.8–99.9 %) in validation studies. The seroprevalence rate of O. lahorensis exposure in serum samples collected from Xinjiang and Inner Mongolia were 10.45 % and 3.09 %, respectively.

Conclusions

The S-iELISA method established in this study was a reliable tool for detecting and monitoring O. lahorensis exposure in sheep. And the serological diagnostic method based on rOl-BTSP showed great promise for enhancing the surveillance and management strategies for O. lahorensis-borne diseases.

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来源期刊

Veterinary parasitology, regional studies and reports Multiple-

CiteScore

2.90

自引率

7.10%

发文量

126

审稿时长

97 days

期刊介绍： Veterinary Parasitology: Regional Studies and Reports focuses on aspects of veterinary parasitology that are of regional concern, which is especially important in this era of climate change and the rapid and often unconstrained travel of people and animals. Relative to regions, this journal will accept papers of the highest quality dealing with all aspects of disease prevention, pathology, treatment, epidemiology, and control of parasites within the field of veterinary medicine. Also, case reports will be considered as they add to information related to local disease and its control; such papers must be concise and represent appropriate medical intervention. Papers on veterinary parasitology from wildlife species are acceptable, but only if they relate to the practice of veterinary medicine. Studies on vector-borne bacterial and viral agents are suitable, but only if the paper deals with vector transmission of these organisms to domesticated animals. Studies dealing with parasite control by means of natural products, both in vivo and in vitro, are more suited for one of the many journals that now specialize in papers of this type. However, due to the regional nature of much of this research, submissions may be considered based upon a case being made by the author(s) to the Editor. Circumstances relating to animal experimentation must meet the International Guiding Principles for Biomedical Research Involving Animals as issued by the Council for International Organizations of Medical Sciences (obtainable from: Executive Secretary C.I.O.M.S., c/o W.H.O., Via Appia, CH-1211 Geneva 27, Switzerland).