Diagnostic efficacy of monoclonal antibody based sandwich enzyme linked immunosorbent assay (ELISA) for detection of Fasciola gigantica excretory/secretory antigens in both serum and stool

Tarek M Diab,Mohy E Attia,Azza E El-Bassiouny,Mona K Zoheiry,Salwa H Mohamed,Zeinab A Demerdash,Faten S Mahmoud,Wafaa A Mansour,Ibrahim R Aly

doi:10.1186/1756-3305-4-176

Tarek M Diab, Mohy E Attia + Show 7 more

Open Access

https://doi.org/10.1186/1756-3305-4-176

Copy DOI

Journal: Parasites & Vectors	Publication Date: Sep 15, 2011
Citations: 57	License type: cc-by

Affiliation: Theodor Bilharz Research Institute

Abstract

BackgroundThis research was carried out to develop a reliable monoclonal antibody (MoAb)-based sandwich enzyme linked immunosorbent assay (ELISA) for the diagnosis of active Fasciola gigantica infection in both serum and stool for comparative purposes.MethodsFrom a panel of MoAbs raised against F. gigantica excretory/secretory antigens (ES Ags), a pair (12B/11D/3F and 10A/9D/10G) was chosen due to its high reactivity and strict specificity to F. gigantica antigen by indirect ELISA.ResultsThe two MoAbs were of the IgG1 and IgG2a subclasses, respectively. Using SDS-PAGE and EITB, the selected MoAbs recognized 83, 64, 45 and 26 kDa bands of ES Ags. The lower detection limit of ELISA assay was 3 ng/ml. In stool, the sensitivity, specificity and diagnostic efficacy of ELISA was 96%, 98.2 and 97.1%; while in serum they were 94%, 94.6% and 94.3%, respectively. Moreover, a positive correlation was found between ova count in stool of F. gigantica infected patients and the OD readings of ELISA in both stool and serum samples (r = 0.730, p < 0.01 and r = 0.608; p < 0.01, respectively).ConclusionsThese data showed that the use of MoAb-based sandwich ELISA for the detection of F. gigantica coproantigens in stool specimens was superior to serum samples; it provides a highly efficient, non-invasive technique for the diagnosis of active F. gigantica infection.

Highlights

This research was carried out to develop a reliable monoclonal antibody (MoAb)-based sandwich enzyme linked immunosorbent assay (ELISA) for the diagnosis of active Fasciola gigantica infection in both serum and stool for comparative purposes
These data showed that the use of MoAb-based sandwich ELISA for the detection of F. gigantica coproantigens in stool specimens was superior to serum samples; it provides a highly efficient, non-invasive technique for the diagnosis of active F. gigantica infection
Fasciola hepatica and F. gigantica are two trematode species which have an important impact on public health due to the infections they cause in humans and livestock

Summary

Introduction

This research was carried out to develop a reliable monoclonal antibody (MoAb)-based sandwich enzyme linked immunosorbent assay (ELISA) for the diagnosis of active Fasciola gigantica infection in both serum and stool for comparative purposes. The majority of cases are attributed to F. hepatica, human infections with F. gigantica are present in many countries [4,5,6]. Detection of circulating Fasciola antigen in both serum and stool was found to be more sensitive and specific [12]. The majority of methods based on antigen detection are applied to F. hepatica infection, but only few are applied to F. gigantica infection [13,14,15]

Methods

Results

Conclusion