A serological test based on mutated recombinant Fasciola hepatica cathepsin L protease for the diagnosis of equine fasciolosis

IF 2.2 2区农林科学 Q2 PARASITOLOGY

Veterinary parasitology Pub Date : 2025-09-02 DOI:10.1016/j.vetpar.2025.110592

Philipp A. Kronenberg , Nathalie Fouché , Mary Sekiya , Pauline Liechti , Caroline F. Frey , Grace Mulcahy , Ramon M. Eichenberger

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引用次数: 0

Abstract

Fasciola hepatica is a common trematode parasite of livestock in many regions, causing significant economic losses and affecting animal welfare. Horses rarely develop patent liver fluke infection. However, liver damage can affect animal health and welfare. Therefore, F. hepatica infection in horses may be underreported. Recently, a serological test for the antibody detection has been reported based on recombinant parasite cathepsin L1 (FhCL1) protease. Here, we optimized this enzyme-linked immunosorbent assay (ELISA) for antibody-detection in horses with equine fasciolosis based on recombinant forms of FhCL proteases with a bioengineered diagnostic epitope mutation. Epitopes from different homologues of FhCL-proteases were modified, and canonical and mutated versions were recombinantly produced. The best performing candidate was evaluated with 175 serum samples from horses in Ireland. Seven horses suspected of having liver fluke infections were monitored for one year following treatment with triclabendazole. Additionally, a cohort of 368 samples from Swiss horses was tested to gather prevalence data. The final diagnostic test based on an epitope-mutated antigen showed a sensitivity and specificity of 65 % and 97.4 %, respectively. Follow-up of horses with suspected F. hepatica infection showed not only improved biochemical liver values but also a decrease in antibody titres, which fell below the test threshold after approximately 6–9 months. This group allowed for the definition of thresholds to distinguish between ambiguous/borderline results and clear positives. The prevalence of equine fasciolosis in Swiss horses was between 3.5 % and 5.7 %, depending on the applied diagnostic threshold. These results suggest that F. hepatica may be a neglected infection in Swiss horses. However, reliable diagnostic tests remain important, particularly for detecting cases in horses with liver disease of unknown origin or those co-grazed with ruminants in endemic regions.

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基于突变重组肝片形吸虫组织蛋白酶L的血清学检测诊断马片形吸虫病

肝片吸虫是许多地区常见的牲畜吸虫，造成重大经济损失并影响动物福利。马很少发生肝吸虫感染。然而，肝脏损伤会影响动物的健康和福利。因此，马的肝杆菌感染可能被低估了。最近，一种基于重组寄生虫组织蛋白酶L1 （FhCL1）蛋白酶的抗体检测血清学方法被报道。在这里，我们优化了这种酶联免疫吸附试验（ELISA），用于马片形虫病的抗体检测，该试验基于具有生物工程诊断表位突变的重组形式的FhCL蛋白酶。对不同同源的fhcl蛋白酶表位进行修饰，重组产生典型和突变版本。用175个爱尔兰马的血清样本对表现最好的候选物进行了评估。7匹疑似有肝吸虫感染的马在使用三氯苯达唑治疗后监测了一年。此外，对来自瑞士马的368个样本进行了队列测试，以收集流行数据。基于表位突变抗原的最终诊断试验的敏感性和特异性分别为65% %和97.4% %。对疑似肝炎F.感染的马进行随访，不仅肝脏生化指标有所改善，而且抗体滴度也有所下降，在大约6-9 个月后降至检测阈值以下。该组允许定义阈值，以区分模糊/边缘性结果和明确阳性结果。根据应用的诊断阈值，瑞士马的马片形吸虫病患病率在3.5% %和5.7% %之间。这些结果表明，瑞士马的肝梭菌可能是一种被忽视的感染。然而，可靠的诊断检测仍然很重要，特别是在发现来历不明的肝病马或在流行地区与反刍动物共食的马的病例时。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Veterinary parasitology 农林科学-寄生虫学

CiteScore

5.30

自引率

7.70%

发文量

126

审稿时长

36 days

期刊介绍： The journal Veterinary Parasitology has an open access mirror journal,Veterinary Parasitology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. This journal is concerned with those aspects of helminthology, protozoology and entomology which are of interest to animal health investigators, veterinary practitioners and others with a special interest in parasitology. Papers of the highest quality dealing with all aspects of disease prevention, pathology, treatment, epidemiology, and control of parasites in all domesticated animals, fall within the scope of the journal. Papers of geographically limited (local) interest which are not of interest to an international audience will not be accepted. Authors who submit papers based on local data will need to indicate why their paper is relevant to a broader readership. Parasitological studies on laboratory animals fall within the scope of the journal only if they provide a reasonably close model of a disease of domestic animals. Additionally the journal will consider papers relating to wildlife species where they may act as disease reservoirs to domestic animals, or as a zoonotic reservoir. Case studies considered to be unique or of specific interest to the journal, will also be considered on occasions at the Editors'' discretion. Papers dealing exclusively with the taxonomy of parasites do not fall within the scope of the journal.