Evaluation of a recombinant cathepsin L1 ELISA and comparison with the Pourquier and ES ELISA for the detection of antibodies against Fasciola hepatica

Kuerpick, Birte; Schnieder, Thomas; Strübe, Christina

doi:10.1016/j.vetpar.2012.11.021

Cited by 30 publications

(36 citation statements)

References 21 publications

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“…Moreover, this phenomenon was not limited to the samples investigated in this study, nor to the expression of rpFhCLs in E. coli. In agreement with this, a similar finding was recently reported in a study where the performance of an indirect ELISA based on a rFhpCL1 expressed in the yeast Pichia pastoris was compared with that of a commercial indirect ELISA test containing a purified fraction from Fasciola ESAs (f2 antigen; Institute Pourquier, Montpellier, France) (Kuerpick et al 2013). Cross-reactivity was also observed by Cornelissen et al (2001) who tested sera from sheep and cattle harboring other parasites, mainly nematodes, suggesting that the cross-reactivity may be due to common epitopes between recombinant Fasciola cathepsins and antigens present in other parasites.…”

Section: Discussionsupporting

confidence: 85%

“…These molecules were selected since (i) they are produced by adult flukes and thus continue stimulating the immune system during the chronic phase of the illness, and (ii) Fasciola cathepsins L1, L2, and L5 contain a common epitope recognized by mAb MM3, which is the capture antibody in the MM3-SERO ELISA (Mezo et al 2007). Although rFhpCLs from the L1 clade have already been successfully used to develop a very sensitive and specific lateral flow test for immunodiagnosis of human fascioliasis , as well as to design ELISA tests for human (O'Neill et al 1999;Carnevale et al 2001a, b;Gonzales Santana et al 2013;Gottstein et al 2014) and animal use (Cornelissen et al 2001;Kuerpick et al 2013;Selemetas et al 2014;Bloemhoff et al 2015), the performance of rFhpCLs from L2 and L5 clades as ELISA targets has not previously been investigated. From a functional point of view, a single amino acid substitution may be sufficient to affect substrate specificity in cysteine proteases from F. hepatica (Smooker et al 2000).…”

Section: Discussionmentioning

confidence: 99%

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Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis

et al. 2018

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Infections caused by Fasciola hepatica are of great importance in the veterinary field, as they cause important economic losses to livestock producers. Serodiagnostic methods, typically ELISA (with either native or recombinant antigens), are often used for early diagnosis. The use of native antigens, as in the MM3-SERO ELISA (commercialized as BIO K 211, BIO-X Diagnostics), continues to be beneficial in terms of sensitivity and specificity; however, there is interest in developing ELISA tests based on recombinant antigens to avoid the need to culture parasites. Of the antigens secreted by adult flukes, recombinant procathepsin L1 (rFhpCL1) is the most commonly tested in ELISA to date. However, although adult flukes produce three different clades of CLs (FhCL1, FhCL2, and FhCL5), to our knowledge, the diagnostic value of recombinant FhCL2 and FhCL5 has not yet been investigated. In the present study, we developed and tested three indirect ELISAs using rFhpCL1, rFhpCL2, and rFhpCL5 and evaluated their recognition by sera from sheep and cattle naturally infected with F. hepatica. Although the overall antibody response to these three rFhpCLs was similar, some animals displayed preferential recognition for particular rFhpCLs. Moreover, for cattle sera, the highest sensitivity was obtained using rFhpCL2 (97%), being equal for both rFhpCL1 and rFhpCL5 (87.9%), after adjusting cut-offs for maximum specificity. By contrast, for sheep sera, the sensitivity was 100% for the three rFhpCLs. Finally, the presence of truncated and/or partially unfolded molecules in antigen preparations is postulated as a possible source of cross-reactivity.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis

et al. 2018

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“…This is different to what has been reported in the literature regarding the use of this antigen. Kuerpick and colleagues [ 22 ] observed two false positive results out of 13 animals infected with D. viviparus and one out of four animals infected with C. oncophora (sensitivity between 90–100 %, specificity 88.6 %). Cornelissen et al .…”

Section: Discussionmentioning

confidence: 99%

“…The generally high handling costs as well as the necessity to sample several animals led to the increased use of serological methods which can be used for herd health monitoring. Serological diagnosis of F. hepatica has been described in the literature using excretory/secretory (ES) products [ 21 – 23 ], a “f2” antigen (Fasciolosis Verification Test, IDEXX, Hoofddorp, the Netherlands) and a recombinant Cathepsin L1 antigen [ 22 ]. The same applies for D. viviparus where the detection of antibodies in serum or milk using ELISAs with either crude ES antigen [ 24 – 26 ] or recombinantly expressed major sperm protein (MSP) [ 27 – 29 ] has been described.…”

Section: Introductionmentioning

confidence: 99%

Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle

et al. 2015

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BackgroundA 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.MethodsThe 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).ResultsEstablishment 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.ConclusionsA 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.

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“…no. ) was linked to a 6x His‐tag, inserted into the expression vector pPinkα‐HC (Invitrogen) and electroporated into the yeast Pichia pastoris as described . PCR with subsequent commercial Sanger sequencing (Seqlab Sequence Laboratories, Göttingen, Germany) verified the correct insert sequence of selected clones.…”

Section: Methodsmentioning

confidence: 99%

Vaccination of calves with yeast‐ and bacterial‐expressed paramyosin from the bovine lungworm Dictyocaulus viviparus

Joekel

Hinse

Raulf

et al. 2015

Parasite Immunology

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Summary Previously, vaccination of cattle with Escherichia coli‐expressed bovine lungworm paramyosin (EcPMY) adjuvanted with Quil A resulted in considerable reduction in worm burden and larvae shedding (Strube et al., 2015). To further evaluate the protective potential of PMY, cattle vaccination trials were performed using either E. coli‐ (EcPMY) or Pichia pastoris‐expressed PMY (PpPMY) with different adjuvants (Matrix‐Q™ or Quil A). Combinations EcPMY+Matrix‐Q™ (trial 1), PpPMY+Matrix‐Q™ (trial 2) and PpPMY+Quil A (trial 3) were tested against challenge infections with 2000 Dictyocaulus viviparus larvae. Even though GM worm burden and larvae shedding was lower in almost all vaccinated groups, there were high variations between individuals hampering significant differences. However, in all vaccinated groups, lungworms were significantly shorter compared with those in controls. In vitro stimulation of peripheral blood mononuclear cells (PBMC) with recombinant (r)PMY revealed no significant proliferation following vaccinations or challenge infection. All vaccinated cattle showed a significant rise in specific antibodies, particularly IgG and its subclass IgG1, and detected the native lungworm PMY in immunoblots starting 2 weeks after the first vaccination. The use of a different rPMY‐adjuvant combination or combined vaccination with additional recombinant antigens might be a promising future approach towards a new vaccine against lungworms in cattle.

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Evaluation of a recombinant cathepsin L1 ELISA and comparison with the Pourquier and ES ELISA for the detection of antibodies against Fasciola hepatica

Cited by 30 publications

References 21 publications

Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis

Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis

Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle

Vaccination of calves with yeast‐ and bacterial‐expressed paramyosin from the bovine lungworm Dictyocaulus viviparus

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