Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle
Abstract Background A major constraint for the effective control and management of helminth parasites is the lack of rapid, high-throughput, routine diagnostic tests to assess the health status of individual animals and herds and to identify the parasite species responsible for these helminthoses. T...
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| Format: | Article |
| Language: | English |
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BMC
2015-06-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13071-015-0924-0 |
| _version_ | 1797811892447608832 |
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| author | Sofia N. Karanikola Jürgen Krücken Sabrina Ramünke Theo de Waal Johan Höglund Johannes Charlier Corinna Weber Elisabeth Müller Slawomir J. Kowalczyk Jaroslaw Kaba Georg von Samson-Himmelstjerna Janina Demeler |
| author_facet | Sofia N. Karanikola Jürgen Krücken Sabrina Ramünke Theo de Waal Johan Höglund Johannes Charlier Corinna Weber Elisabeth Müller Slawomir J. Kowalczyk Jaroslaw Kaba Georg von Samson-Himmelstjerna Janina Demeler |
| author_sort | Sofia N. Karanikola |
| collection | DOAJ |
| description | Abstract Background A major constraint for the effective control and management of helminth parasites is the lack of rapid, high-throughput, routine diagnostic tests to assess the health status of individual animals and herds and to identify the parasite species responsible for these helminthoses. The capability of a multiplex platform for the simultaneous detection of three pasture associated parasite species was evaluated and compared to existing ELISAs. Methods The recombinant antigens 14.2 kDa ES protein for Cooperia oncophora, major sperm protein for Dictyocaulus viviparus and Cathepsin L1 for Fasciola hepatica were recombinantly expressed either in Escherichia coli or Pichia pastoris. Antigens were covalently coupled onto magnetic beads. Optimal concentrations for coupling were determined following the examination of serum samples collected from experimentally mono-infected animals, before and after their infection with the target species. Absence of cross-reactivity was further determined with sera from calves mono-infected with Haemonchus contortus, Ostertagia ostertagi and Trichostrongylus colubriformis. Examination of negative serum samples was characterised by low median fluorescence intensity (MFI). Results Establishment of the optimal serum dilution of 1:200 was achieved for all three bead sets. Receiver Operating Characteristic analyses were performed to obtain cut-off MFI values for each parasite separately. Sensitivity and specificity at the chosen cut-off values were close to, or 100 % for all bead sets. Examination of serum samples collected on different days post infection from different animals showed a high reproducibility of the assays. Serum samples were additionally examined with two already established ELISAs, an in-house ELISA using the recombinant MSP as an antigen and a DRG ELISA using Cathepsin L1 for liver fluke. The results between the assays were compared and kappa tests revealed an overall good agreement. Conclusions A versatile bead-based assay using fluorescence detection (xMAP® technology) was developed to simultaneously detect antibodies against C. oncophora, D. viviparus and F. hepatica in cattle serum samples. This platform provides rapid, high-throughput results and is highly sensitive and specific in comparison to existing serological as well as coproscopical diagnostic techniques. |
| first_indexed | 2024-03-13T07:30:29Z |
| format | Article |
| id | doaj.art-642e59a51bc741cc8ea3441be7134082 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-03-13T07:30:29Z |
| publishDate | 2015-06-01 |
| publisher | BMC |
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| series | Parasites & Vectors |
| spelling | doaj.art-642e59a51bc741cc8ea3441be71340822023-06-04T11:09:20ZengBMCParasites & Vectors1756-33052015-06-018111110.1186/s13071-015-0924-0Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattleSofia N. Karanikola0Jürgen Krücken1Sabrina Ramünke2Theo de Waal3Johan Höglund4Johannes Charlier5Corinna Weber6Elisabeth Müller7Slawomir J. Kowalczyk8Jaroslaw Kaba9Georg von Samson-Himmelstjerna10Janina Demeler11Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität BerlinInstitute for Parasitology and Tropical Veterinary Medicine, Freie Universität BerlinInstitute for Parasitology and Tropical Veterinary Medicine, Freie Universität BerlinUCD School of Agriculture, Food Science and Veterinary Medicine, University College DublinDepartment of Biomedical Sciences and Veterinary Public Health, Section for Parasitology (SWEPAR), Swedish University of Agricultural SciencesDepartment of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent UniversityLABOKLIN GMBH & Co.KGLABOKLIN GMBH & Co.KGLaboratory of Veterinary Epidemiology and Economics, Faculty of Veterinary Medicine, Warsaw University of Life ScienceLaboratory of Veterinary Epidemiology and Economics, Faculty of Veterinary Medicine, Warsaw University of Life ScienceInstitute for Parasitology and Tropical Veterinary Medicine, Freie Universität BerlinInstitute for Parasitology and Tropical Veterinary Medicine, Freie Universität BerlinAbstract Background A major constraint for the effective control and management of helminth parasites is the lack of rapid, high-throughput, routine diagnostic tests to assess the health status of individual animals and herds and to identify the parasite species responsible for these helminthoses. The capability of a multiplex platform for the simultaneous detection of three pasture associated parasite species was evaluated and compared to existing ELISAs. Methods The recombinant antigens 14.2 kDa ES protein for Cooperia oncophora, major sperm protein for Dictyocaulus viviparus and Cathepsin L1 for Fasciola hepatica were recombinantly expressed either in Escherichia coli or Pichia pastoris. Antigens were covalently coupled onto magnetic beads. Optimal concentrations for coupling were determined following the examination of serum samples collected from experimentally mono-infected animals, before and after their infection with the target species. Absence of cross-reactivity was further determined with sera from calves mono-infected with Haemonchus contortus, Ostertagia ostertagi and Trichostrongylus colubriformis. Examination of negative serum samples was characterised by low median fluorescence intensity (MFI). Results Establishment of the optimal serum dilution of 1:200 was achieved for all three bead sets. Receiver Operating Characteristic analyses were performed to obtain cut-off MFI values for each parasite separately. Sensitivity and specificity at the chosen cut-off values were close to, or 100 % for all bead sets. Examination of serum samples collected on different days post infection from different animals showed a high reproducibility of the assays. Serum samples were additionally examined with two already established ELISAs, an in-house ELISA using the recombinant MSP as an antigen and a DRG ELISA using Cathepsin L1 for liver fluke. The results between the assays were compared and kappa tests revealed an overall good agreement. Conclusions A versatile bead-based assay using fluorescence detection (xMAP® technology) was developed to simultaneously detect antibodies against C. oncophora, D. viviparus and F. hepatica in cattle serum samples. This platform provides rapid, high-throughput results and is highly sensitive and specific in comparison to existing serological as well as coproscopical diagnostic techniques.https://doi.org/10.1186/s13071-015-0924-0Parasitic gastroenteritisLungwormLiver flukeLuminexMultiplex immunoassaysSerum |
| spellingShingle | Sofia N. Karanikola Jürgen Krücken Sabrina Ramünke Theo de Waal Johan Höglund Johannes Charlier Corinna Weber Elisabeth Müller Slawomir J. Kowalczyk Jaroslaw Kaba Georg von Samson-Himmelstjerna Janina Demeler Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle Parasites & Vectors Parasitic gastroenteritis Lungworm Liver fluke Luminex Multiplex immunoassays Serum |
| title | Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle |
| title_full | Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle |
| title_fullStr | Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle |
| title_full_unstemmed | Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle |
| title_short | Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle |
| title_sort | development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against cooperia oncophora dictyocaulus viviparus and fasciola hepatica in cattle |
| topic | Parasitic gastroenteritis Lungworm Liver fluke Luminex Multiplex immunoassays Serum |
| url | https://doi.org/10.1186/s13071-015-0924-0 |
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