Diagnosis of Fasciola gigantica infection using a monoclonal antibody-based sandwich ELISA for detection of circulating cathepsin B3 protease

Anuracpreeda, Panat; Chawengkirtikul, Runglawan; Tinikul, Yotsawan; Poljaroen, Jaruwan; Chotwiwatthanakun, Charoonroj; Sobhon, Prasert

doi:10.1016/j.actatropica.2013.03.020

Cited by 30 publications

(32 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the earlier reports, ELISA and proteomic assays based on WB antigens of Fasciola sp. have been demonstrated as potential diagnostic tools for immunodiagnosis of Fasciola infections in humans and animals . Therefore, it is possible that this 16 kDa protein could be used as a candidate antigen for the immunodiagnosis of cattle paramphistomosis caused by P. gracile , but would require further development.…”

Section: Discussionmentioning

confidence: 99%

Antigenic profile, isolation and characterization of whole body extract of Paramphistomum gracile

Anuracpreeda

Chawengkirttikul

Sobhon

2016

Parasite Immunology

View full text Add to dashboard Cite

An antigenic component of adult Paramphistomum gracile was characterized by means of indirect enzyme-linked immunosorbent assay (indirect ELISA), sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using sera from cattle naturally infected with P. gracile, Eurytrema pancreaticum, Fasciola gigantica, Moniezia benedeni, strongylids, Trichuris sp. and Strongyloides sp. The whole body (WB) extracts of P. gracile were fractionated by gel filtration chromatography in a Sephadex G-200 column. It was found that the WB extract fractions, F1-F3 were highly antigenic, F5 was moderately antigenic and F4 was poorly antigenic. For SDS-PAGE and immunoblotting, the antigenic molecules of WB extract and all five fractions were mostly at molecular weights (MW) ranging from 12 to 150 kDa. One antigenic protein of 16 kDa detected in WB extract and F1-F3 was found to give a consistent reaction with sera from infected cattle. The antigenicity of the purified 16 kDa protein was confirmed by immunoblotting and indirect ELISA using a pool of sera and individual serum samples from infected cattle (at 1 : 78 125 dilution) and hyperimmunized rabbit (at 1 : 390 625 dilution). This finding suggests that the 16 kDa protein may be a potential antigen for the immunodiagnosis of cattle paramphistomosis caused by P. gracile.

show abstract

Section: Discussionmentioning

confidence: 99%

Antigenic profile, isolation and characterization of whole body extract of Paramphistomum gracile

Anuracpreeda

Chawengkirttikul

Sobhon

2016

Parasite Immunology

View full text Add to dashboard Cite

show abstract

“…The flukes were washed several times with 0.85% NaCl solution and processed immediately for SEM and LM according to the methods described by Panyarachun et al [20] and Anuracpreeda et al [1,3]. …”

Section: Methodsmentioning

confidence: 99%

Morphology and histology of the adultParamphistomum gracileFischoeder, 1901

et al. 2013

Self Cite

View full text Add to dashboard Cite

In the present study, we evaluated the histological morphology of the adult Paramphistomum (P.) gracile. Adult flukes with bodies 5~15 mm in length and 2~7 mm in width were subjected to histological analysis. Longitudinal and transversal serial-sections were stained with hematoxylin and eosin, and examined. The body surface and longitudinal section of P. gracile were also assessed using scanning electron microscopy. In this species, the anterior sucker and posterior sucker (acetabulum) were present on an anterior and posterior part of the body, respectively. The major folds were located in the areas of the anterior sucker, genital canal, and posterior sucker. The fluke membrane was spineless at the tegument surface and in the tegument tissue. Histological data showed structural-systematic characteristics of the digestive tract, reproductive tract, excretory tract, copulatory organs, connective tissues, and muscle tissues. We attempted to elucidate the histological characteristics of P. gracile that might increase the knowledge and understanding of rumen fluke morphology.

show abstract

“…Therefore, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden. Only a few detections of circulating antigens in animals with fasciolosis have been reported with variable efficiencies due largely to their availabilities in the circulation [ 8 – 13 ].…”

Section: Introductionmentioning

confidence: 99%

Immunodiagnosis of Fasciola gigantica Infection Using Monoclonal Antibody-Based Sandwich ELISA and Immunochromatographic Assay for Detection of Circulating Cathepsin L1 Protease

2016

Self Cite

View full text Add to dashboard Cite

BackgroundTropical fasciolosis caused by Fasciola gigantica infection is one of the major diseases infecting ruminants in the tropical regions of Africa and Asia including Thailand. Parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Therefore, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden.MethodsIn this study, we have produced a monoclonal antibody (MoAb) against recombinant F. gigantica cathepsin L1 (rFgCatL1), and developed both sandwich enzyme-linked immunosorbent assay (sandwich ELISA) and immunochromatographic (IC) test for rapid detection of circulating cathepsin L1 protease (CatL1) in the sera from mice experimentally and cattle naturally infected with Fasciola gigantica. MoAb 4E3 and biotinylated rabbit anti-recombinant CatL1 antibody were selected due to their high reactivities and specificities.ResultsThe lower detection limits of sandwich ELISA and IC test were 3 pg/ml and 0.256 ng/ml, respectively. Sandwich ELISA and IC test could detect F. gigantica infection from day 1 to 35 post infection. In experimental mice, the sensitivity, specificity and accuracy were 95%, 100% and 98.6% (for sandwich ELISA), and 93%, 100% and 98.2% (for IC test), while in natural cattle they were 98.3%, 100% and 99.5% (for sandwich ELISA), and 96.7%, 100% and 99.1% (for IC test).ConclusionsThese two assay methods showed high efficiencies and precisions for diagnosis of fasciolosis by F. gigantica.

show abstract

Diagnosis of Fasciola gigantica infection using a monoclonal antibody-based sandwich ELISA for detection of circulating cathepsin B3 protease

Cited by 30 publications

References 26 publications

Antigenic profile, isolation and characterization of whole body extract of Paramphistomum gracile

Antigenic profile, isolation and characterization of whole body extract of Paramphistomum gracile

Morphology and histology of the adultParamphistomum gracileFischoeder, 1901

Immunodiagnosis of Fasciola gigantica Infection Using Monoclonal Antibody-Based Sandwich ELISA and Immunochromatographic Assay for Detection of Circulating Cathepsin L1 Protease

Contact Info

Product

Resources

About