Philipp A. Kronenberg , Nathalie Fouché , Mary Sekiya , Pauline Liechti , Caroline F. Frey , Grace Mulcahy , Ramon M. Eichenberger
{"title":"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","authors":"Philipp A. Kronenberg , Nathalie Fouché , Mary Sekiya , Pauline Liechti , Caroline F. Frey , Grace Mulcahy , Ramon M. Eichenberger","doi":"10.1016/j.vetpar.2025.110592","DOIUrl":null,"url":null,"abstract":"<div><div><em>Fasciola hepatica</em> 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, <em>F. hepatica</em> 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 <em>F. hepatica</em> 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 <em>F. hepatica</em> 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.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110592"},"PeriodicalIF":2.2000,"publicationDate":"2025-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"A serological test based on mutated recombinant Fasciola hepatica cathepsin L protease for the diagnosis of equine fasciolosis\",\"authors\":\"Philipp A. Kronenberg , Nathalie Fouché , Mary Sekiya , Pauline Liechti , Caroline F. Frey , Grace Mulcahy , Ramon M. Eichenberger\",\"doi\":\"10.1016/j.vetpar.2025.110592\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div><em>Fasciola hepatica</em> 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, <em>F. hepatica</em> 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 <em>F. hepatica</em> 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 <em>F. hepatica</em> 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.</div></div>\",\"PeriodicalId\":23716,\"journal\":{\"name\":\"Veterinary parasitology\",\"volume\":\"340 \",\"pages\":\"Article 110592\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2025-09-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Veterinary parasitology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0304401725002031\",\"RegionNum\":2,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"PARASITOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Veterinary parasitology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0304401725002031","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PARASITOLOGY","Score":null,"Total":0}
A serological test based on mutated recombinant Fasciola hepatica cathepsin L protease for the diagnosis of equine fasciolosis
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.
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.