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

Panat Anuracpreeda,Prasert Sobhon,Runglawan Chawengkirttikul,Massimiliano Galdiero

doi:10.1371/journal.pone.0145650

Panat Anuracpreeda, Prasert Sobhon + Show 2 more

Open Access

https://doi.org/10.1371/journal.pone.0145650

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Journal: PloS one	Publication Date: Jan 5, 2016
Citations: 28	License type: CC BY 4.0

Affiliation: Mahidol University, Kanchanaburi Rajabhat University

Abstract

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.

Highlights

Tropical fasciolosis caused by Fasciola gigantica infection is one of the major diseases infecting ruminants in the tropical regions of Asia and Africa, and causes a significant economic loss in livestock industry in developing and underdeveloped countries for more than 3.2 billion US dollars per annum [1]
We previously demonstrated that the full-length FgCatL1 cDNA was successfully subcloned into the bacterial expression vector, pET-30b (Novagen), which was transformed into Escherichia coli BL21 (DE)
monoclonal antibody (MoAb) 4E3 showed the highest optical density (OD) values of antibody reactivities against rFgCatL1 and the native FgCatL1 in Crude extract (CE) and ES fractions when screened by indirect ELISA; it has been used in the present study

Summary

Introduction

Tropical fasciolosis caused by Fasciola gigantica infection is one of the major diseases infecting ruminants in the tropical regions of Asia and Africa, and causes a significant economic loss in livestock industry in developing and underdeveloped countries for more than 3.2 billion US dollars per annum [1]. The serodiagnostic method for the detection of antibody in animals has been developed for the diagnosis of fasciolosis [5,6,7]; the circulating antibodies appear in the serum for several months after the infection even though the parasites may have already been killed. Tropical fasciolosis caused by Fasciola gigantica infection is one of the major diseases infecting ruminants in the tropical regions of Africa and Asia including Thailand. The detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden.

Methods

Results

Conclusion