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

Demerdash, Zeinab; Diab, T.; Aly, Ibrahim; Mohamed, Salwa H.; Mahmoud, Faten S.; Zoheiry, Mona; Mansour, Wafaa; Attia, Mohy; El-Bassiouny, A.

doi:10.1186/1756-3305-4-176

Cited by 35 publications

(30 citation statements)

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“…Interestingly, our developed assay shows results that are superior or at least similar to those of the previously reported antigen tests for diagnosis of human fascioliasis (37,51,52), and the differences may be due to the different natures of antigens used. A recent study evaluated an antigen capture ELISA using a pair of monoclonal antibodies raised against F. gigantica E/S antigens in human serum and stools at a lower detection limit of 3 ng/ml (25). The sensitivity (94%) and specificity (94.6%) of the capture ELISA in serum were similar to those shown in our study.…”

Section: Discussionsupporting

confidence: 70%

“…Several sensitive and specific methods based on Fasciola antigen detection in serum and stool using specific antibodies were applied to F. hepatica and F. gigantica infections in animals (8,25,26,47). The antigen tests can detect experimental infection a few days after inoculation (23).…”

Section: Discussionmentioning

confidence: 99%

“…In that sense, the direct detection of parasite antigens in stool (coproantigens) or serum (circulating antigens) of Fasciola-infected animals or humans has been reported as a new alternative approach with greater diagnostic accuracy (22)(23)(24)(25)(26). In our previous study, we identified a 26-to 28-kDa circulating antigen in sera of cattle infected with F. gigantica by using specific rabbit IgG antiserum (22).…”

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confidence: 99%

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Immunodetection of Fasciola gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis

Attallah¹,

Bughdadi

El-Shazly

et al. 2013

Clin. Vaccine Immunol.

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c Currently, the laboratory diagnosis of human fascioliasis is based on the parasitological examination of parasite eggs in stool specimens and serological detection of specific antibodies in serum samples, which are often unreliable diagnostic approaches. Ideally, a sensitive and specific diagnostic test for Fasciola infection should be based on the detection of circulating Fasciola antigen, which implies active infection. Here, a 27-kDa-molecular-mass antigen was identified in a Fasciola gigantica adult worm antigen preparation, excretory-secretory products, and sera from F. gigantica-infected individuals, and it was not detected in antigenic extracts of other parasites and sera from noninfected individuals. The target antigen was isolated and partially characterized as a protein. Immunoperoxidase staining located the target epitope within teguments and guts of F. gigantica adult worms. The performance characteristics of a newly developed enzyme-linked immunosorbent assay (ELISA) based on F. gigantica circulating antigen detection in serum (FgCA-27 ELISA) were investigated using sera of 120 parasitologically diagnosed F. gigantica-infected individuals and 80 noninfected individuals. The area under the receiving operating characteristic (ROC) curve (AUC) for ELISA was significantly high (AUC ‫؍‬ 0.961, P < 0.0001) for discriminating Fasciola-infected and noninfected individuals. The developed assay showed high degrees of sensitivity, specificity, and efficiency (>93%), and a significant correlation (r ‫؍‬ 0.715, P < 0.0001) between antigen level and parasite egg count was shown. In conclusion, a 27-kDa Fasciola antigen was identified in sera of F. gigantica-infected individuals. A highly sensitive and specific Fasciola antigen detection assay, FgCA-27 ELISA, was developed for laboratory diagnosis of human fascioliasis.

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Section: Discussionsupporting

confidence: 70%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Immunodetection of Fasciola gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis

Attallah¹,

Bughdadi

El-Shazly

et al. 2013

Clin. Vaccine Immunol.

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“…A number of diagnostic tools have been developed for single or simultaneous detection/discrimination of F. hepatica and F. gigantica, including morphological (using shape and size and morphometric features), immunodiagnostic (using monoclonal antibodies or copro-antigen (extracted from eggs in feces) and metacercarial/Fasciola-specific antigens for ELISA) and DNA-related loop-mediated isothermal amplification (LAMP), PCR (single or multiplex), restriction enzymatic, and sequencing methods (2,3,5,11,12,13,35). All the diagnostic methods developed so far have contributed to fast, accurate, and specific detection of Fasciola spp.…”

Section: Discussionmentioning

confidence: 99%

“…To date, various diagnostic techniques for the identification and discrimination of Fasciola spp. have been developed, including the traditional detection of eggs in feces directly or after Kato-Katz based sedimentation (19,43), the morphological examination of adults (38), the lateral flow immunoassay for serodiagnosis (28), and the enzyme-linked immunosorbent assay (indirect ELISA) (12,13,32,36).…”

mentioning

confidence: 99%

Development and Evaluation of a Single-Step Duplex PCR for Simultaneous Detection of Fasciola hepatica and Fasciola gigantica (Family Fasciolidae, Class Trematoda, Phylum Platyhelminthes)

Nguyen

et al. 2012

J Clin Microbiol

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A single-step multiplex PCR (here referred to as a duplex PCR) has been developed for simultaneous detection and diagnosis of Fasciola hepatica and F. gigantica. These species overlap in distribution in many countries of North and East Africa and Central and Southeast Asia and are similar in egg morphology, making identification from fecal samples difficult. Based on a comparative alignment of mitochondrial DNA (mtDNA) spanning the region of cox1-trnT-rrnL, two species-specific forward primers were designed, FHF (for F. hepatica) and FGF (for F. gigantica), and a single reverse primer, FHGR (common for both species). Conventional PCR followed by sequencing was applied using species-specific primer pairs to verify the specificity of primers and the identity of Fasciola DNA templates. Duplex PCR (using three primers) was used for testing with the DNA extracted from adult worms, miracidia, and eggs, producing amplicons of 1,031 bp for F. hepatica and 615 bp for F. gigantica. The duplex PCR failed to amplify from DNA of other common liver and intestinal trematodes, including two opisthorchiids, three heterophyids, an echinostomid, another fasciolid, and a taeniid cestode. The sensitivity assay showed that the duplex PCR limit of detection for each Fasciola species was between 0.012 ng and 0.006 ng DNA. Evaluation using DNA templates from 32 Fasciola samples (28 adults and 4 eggs) and from 25 field-collected stools of ruminants and humans revealed specific bands of the correct size and the presence of Fasciola species. This novel mtDNA duplex PCR is a sensitive and fast tool for accurate identification of Fasciola species in areas of distributional and zonal overlap.

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Monoclonal Antibodies: Diagnostic Uses

Zola¹,

Mohandas²,

Krumbiegel³

2013

Encyclopedia of Life Sciences

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Diagnosis of disease is a complex process requiring the clinician to deduce, from a set of symptoms and observations, an underlying cause, to predict which of the several therapeutic options is most likely to be effective, and to monitor that effectiveness. Laboratory tests that identify infectious agents, abnormal cells or elements of the body's response to disease, that quantify or localise particular molecules, cells or organisms in body fluids or tissues can provide valuable information to help the clinician with the initial diagnosis, with differential diagnosis (when the symptoms are consistent with several alternative causes) and with selection and monitoring of therapy. Antibodies, because of their exquisite specificity, are particularly useful reagents in this context, and monoclonal antibodies generally show superior specificity compared with polyclonal mixtures of antibodies. Monoclonal antibodies are used widely in the diagnosis of disease, whether in the diagnostic laboratory, in the doctor's rooms or in the field. Key Concepts: Correct diagnosis is a prerequisite to appropriate treatment of disease. Antibodies make useful reagents for identifying infectious or other disease‐causing agents. Monoclonal antibodies are particularly specific and often provide the best diagnostic reagents. Antibodies can be used in a variety of assay formats, designed to answer different questions – presence or absence of a particular substance, amount present and localisation within tissues. The rapid identification of new or rare infectious agents is an important public health measure, to monitor and reduce the chances of epidemics (as we see every few years with a new threat from an influenza variant). Disease often results from imbalances or defects in normal physiological mechanisms, especially the immune system. Many diagnostic tests therefore analyse the components of normal physiological processes. Laboratory tests are useful after the initial diagnosis, to monitor the effects of therapy. Diagnostic tests are generally done in a specialised diagnostic laboratory, but it is often advantageous to perform tests at ‘point of care’ or at home.

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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

Cited by 35 publications

References 33 publications

Immunodetection of Fasciola gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis

Immunodetection of Fasciola gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis

Development and Evaluation of a Single-Step Duplex PCR for Simultaneous Detection of Fasciola hepatica and Fasciola gigantica (Family Fasciolidae, Class Trematoda, Phylum Platyhelminthes)

Monoclonal Antibodies: Diagnostic Uses

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