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使用粗制和重组纤连蛋白抗原对实验感染犬细粒棘球绦虫进行血清学诊断及治疗后监测

Serological diagnosis and post-treatment monitoring of Echinococcus granulosus in experimentally infected dogs using crude and recombinant fibronectin antigens.

作者信息

Celik Figen, Selcuk Muhammed Ahmed, Uslug Muhammet, Simsek Sami

机构信息

Firat University, Faculty of Veterinary Medicine, Department of Parasitology, Elazig, Turkiye.

Firat University, Faculty of Veterinary Medicine, Department of Parasitology, Elazig, Turkiye; Siirt University, Faculty of Veterinary Medicine, Department of Parasitology, Siirt, Turkiye.

出版信息

Vet Immunol Immunopathol. 2025 Jul;285:110962. doi: 10.1016/j.vetimm.2025.110962. Epub 2025 May 31.

Abstract

Echinococcus granulosus is a zoonotic helminth responsible for cystic echinococcosis, a significant public health concern. The diagnosis of E. granulosus infections in definitive hosts, such as dogs, is challenging due to the absence of clinical signs. This study aimed to evaluate the diagnostic performance of crude (EgSCA) and recombinant (rEgFN162) antigens for the detection of E. granulosus infection in dogs using ELISA and Western blot assays. Additionally, it sought to identify the most suitable antigen and method for population-based screening and post-treatment monitoring. Adult E. granulosus parasites were collected from experimentally infected dogs using arecoline hydrobromide purgation. Soluble crude antigen (EgSCA) was prepared through freeze-thaw cycles, sonication, and filtration, while recombinant fibronectin protein (rEgFN162) was obtained via gene cloning, expression, and purification in E. coli. The antigenic properties of EgSCA and rEgFN162 were analyzed by SDS-PAGE and Western blot. ELISA assays were performed to assess IgG and IgM responses in experimentally infected and treated dogs. Based on IgG ELISA results, EgSCA showed a sensitivity of 96.66 % and specificity of 66.66 %, while rEgFN162 demonstrated a sensitivity of 76.66 % and specificity of 46.66 %. In Western blot analysis, EgSCA achieved a sensitivity of 90 % and specificity of 83.33 %, whereas rEgFN162 showed 66.66 % sensitivity and 73.33 % specificity. The recombinant antigen showed a higher ability to differentiate E. granulosus infections from other helminth infections. The findings suggest that rEgFN162 is a promising candidate for the serodiagnosis of E. granulosus in dogs, with potential applications in epidemiological studies and post-treatment follow-up. Further validation with larger sample sizes is needed to confirm its diagnostic accuracy in natural infections.

摘要

细粒棘球绦虫是一种人畜共患的蠕虫,可引发囊型棘球蚴病,这是一个重大的公共卫生问题。由于缺乏临床症状,在终末宿主(如犬类)中诊断细粒棘球绦虫感染具有挑战性。本研究旨在评估粗抗原(EgSCA)和重组抗原(rEgFN162)通过酶联免疫吸附测定(ELISA)和蛋白质印迹法检测犬类细粒棘球绦虫感染的诊断性能。此外,研究还试图确定用于基于群体的筛查和治疗后监测的最合适抗原和方法。使用氢溴酸槟榔碱泻下法从实验感染的犬类中收集成年细粒棘球绦虫寄生虫。通过冻融循环、超声处理和过滤制备可溶性粗抗原(EgSCA),而重组纤连蛋白蛋白(rEgFN162)则通过基因克隆、表达并在大肠杆菌中纯化获得。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)和蛋白质印迹法分析EgSCA和rEgFN162的抗原特性。进行ELISA检测以评估实验感染和治疗犬类中的IgG和IgM反应。基于IgG ELISA结果,EgSCA的灵敏度为96.66%,特异性为66.66%,而rEgFN162的灵敏度为76.66%,特异性为46.66%。在蛋白质印迹分析中,EgSCA的灵敏度为90%,特异性为83.33%,而rEgFN162的灵敏度为66.66%,特异性为73.33%。重组抗原在区分细粒棘球绦虫感染与其他蠕虫感染方面表现出更高的能力。研究结果表明,rEgFN162是犬类细粒棘球绦虫血清学诊断的一个有前景的候选者,并在流行病学研究和治疗后随访中具有潜在应用价值。需要更大样本量的进一步验证以确认其在自然感染中的诊断准确性。

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