Dot-Blot Immunoassay of Fasciola gigantica Infection using 27 kDa and Adult Worm Regurge Antigens in Egyptian Patients

Kamel, Hanan Hussein; Sarhan, Rania M.

doi:10.3347/kjp.2013.51.2.177

Cited by 12 publications

(9 citation statements)

References 19 publications

(23 reference statements)

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“…Superiority of Fasciola Ag fractions in the range of (25-32) kDa in accurate diagnosis was proved previously by several authors as Farghaly et al [1] for (23-28 kDa) [23] and [8] for 27 kDa using the EITB technique.…”

Section: Discussionsupporting

confidence: 63%

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Diagnosis ofFasciolainfection by SDS–PAGE eluted excretory secretory (ES) protein fractions using dot-ELISA

Sabry

Taher

Allah

et al. 2014

International Journal of Veterinary Science and Medicine

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Fascioliasis is now recognized as an emerging zoonotic disease in Egypt. Diagnosis in suspected patients still needs some degree of accuracy. In the present study, three Fasciola gigantica execratory secretory (ES) protein bands of molecular weight (MW) ranging from 14 to 20 kDa, 25 to 32 kDa and 45 to 65 kDa were eluted after fractionation of the parasite antigen using SDS-PAGE. The extracted kDa protein bands were concentrated and evaluated in diagnosis of Fasciola infection. Moreover the level of their cross reaction with other parasitic infections in infected and suspected patients of known parasite eggs/gram stool was evaluated using the dot-ELISA technique. Protein bands in the range of 14-20 kDa and that of 25-32 kDa were markedly specific and sensitive in diagnosis of different levels of anti-Fasciola antibodies (Ab) in sera of infected cases. These two groups of bands were able to exclude cross-reaction between anti-Fasciola Ab and other parasites recorded in stool of selected patients suffering from Schistosoma mansoni, Ascaris, and Giardia, either in single or mixed conditions with Fasciola eggs. While that of 45-65 kDa appeared less specific than the other previously mentioned bands. Protein bands in the range of 25-32 kDa appeared more sensitive than the other protein bands in detection of anti-Fasciola Ab at higher serum dilutions. The Dot-ELISA technique was proved to be more economic and easy in application. The dotted very small amount of antigens can be stored in a freezer and used at request in diagnosis of large numbers of samples. ª 2014 Production and hosting by Elsevier B.V. on behalf of Faculty of Veterinary Medicine, Cairo University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/ licenses/by-nc-nd/3.0/).

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Section: Discussionsupporting

confidence: 63%

“…At the same time Kamel et al [8] determined that the most specific protein band in F. hepatica ES Ag is that of 14-38 kDa band and the most specific one is that of 27 kDa.…”

Section: Introductionmentioning

confidence: 98%

Diagnosis ofFasciolainfection by SDS–PAGE eluted excretory secretory (ES) protein fractions using dot-ELISA

Sabry

Taher

Allah

et al. 2014

International Journal of Veterinary Science and Medicine

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“…A number of other studies also detected a 27 kDa protein band as an immunodominant band of both somatic and E/S of Fasciola species (Santiago and Hillyer 1988; Attallah et al, 2002; Dixit et al, 2008; Kamel et al, 2013). However the studies of Sampaio Silva et al, (1996), Hammami et al, (1997) and Rokni et al, (2004), revealed the protein band of 29 kDa molecular weight as the principle band of the E/S antigen of Fasciola species for the serodiagnosis of fasciolosis.…”

Section: Discussionmentioning

confidence: 82%

Molecular characterization and Immunodiagnostic potential of various antigenic proteins of Fasciola gigantica species isolated from sheep of North West Himalayan Region

et al. 2019

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SummaryThe control of the digenetic trematode Fasciola gigantica has been the major challenge in both cattle and small ruminants as there is a paucity of an effective and commercial vaccine. Thus, the accurate identification and prepatent diagnosis of F. gigantica is an essential prerequisite for its successful prevention and control. In the present study, the morphologically identified specimens isolated from the liver and bile ducts of sheep (Ovis aries) were validated through molecular data. The sequence analysis of ITS2 of our isolates showed high degree of similarity with F. gigantica and F. hepatica using BLAST function of NCBI. The phylogenetic analysis of our isolates showed a close relationship with previously described F. gigantica and F. hepatica isolates from different countries. The antigenic profile of somatic and E/S antigens of F. gigantica were revealed by SDS–PAGE and immunoblotting using sera from sheep naturally infected with F. gigantica. By SDS-PAGE, 20 distinct bands were revealed from crude somatic fraction. Immunoblotting analysis of these proteins with positive sera exhibited 8 sero-reactive bands ranging from 14 to 97 kDa. Among these 38 and 44 kDa bands were quite specific with high diagnostic specificity and sensitivity. The E/S fraction comprised 7 distinct bands, as revealed by SDS-PAGE analysis. Immunoblotting analysis of these proteins with positive sera exhibited 6 antigenic bands ranging from 23 – 54 kDa. Among these 27 and 33 kDa were found to be quite specific with high diagnostic specificity and sensitivity. The present study concludes that the protein bands of 38 and 44 kDa in somatic fraction and 27 and 33 kDa in E/S fraction can be used for the immunodiagnostic purpose for this economically important parasite, which may also entice further studies regarding their vaccine potential.

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“…The 27 kDa polypeptide of F . gigantica has been studied by previous investigators in the dot blot immunoassay for the detection of fasciolosis in Egyptian patients [ 60 ]. In another study the 27 kDa has been used for the immunodiagnosis of human fasciolosis [ 61 ].…”

Section: Discussionmentioning

confidence: 99%

Immunolocalization and immunodetection of the excretory/secretory (ES) antigens of Fasciola gigantica

Khan

Rehman

et al. 2017

PLoS ONE

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The digenetic trematode Fasciola gigantica is a parasite of great agricultural and economic importance. Along with Fasciola hepatica, F. gigantica incurs huge economic losses to the agricultural sector. Because of unavailability of an effective and commercial vaccine, the earliest diagnosis of the disease is the only way to control the disease. The conventional coprological techniques are able to detect the disease only after the parasites get matured and starts releasing their eggs with the faeces of host, therefore prepatent infection remain undiagnosed. The alternative method is by serological tests that uses circulatory antigens. Despite high sensitivity, their reliability is quite low because of the common antigens shared between different helminth parasites. To overcome this, investigation was shifted to identify the copro-antigens which could be more sensitive and reliable. In the present study, we tried to identify some of the immunodominant proteins from the Excretory Secretory (ES) product of F. gigantica which can be further characterized and used for early detection of infection and also as drug and vaccine candidates. The ES products of F. gigantica were collected and used for raising the polyclonal antibody in rabbit. The polypeptide profile was generated as well as immunogenic polypeptides were identified. The Source of ES antigen was immunolocalized using confocal microscopy and dot blot assay was performed to diagnose field infection. The polypeptide profile of ES products revealed a total of 24 polypeptides out of which 12 immunogenic polypeptides were identified by western blotting. Confocal micrographs showed the immunolocalization of antigens in the intestinal caecae, vitalline glands, gonads as well as in the tegument of the worm. The dot blot assay confirmed the utility of ES products for the detection of field infection. Subsequently, cross reactivity was found negative with Gigantocotyle explanatum; an amphitome parasite of same habitat. However, the cross reactivity with other helminths needs to be worked out.

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Dot-Blot Immunoassay of Fasciola gigantica Infection using 27 kDa and Adult Worm Regurge Antigens in Egyptian Patients

Cited by 12 publications

References 19 publications

Diagnosis ofFasciolainfection by SDS–PAGE eluted excretory secretory (ES) protein fractions using dot-ELISA

Diagnosis ofFasciolainfection by SDS–PAGE eluted excretory secretory (ES) protein fractions using dot-ELISA

Molecular characterization and Immunodiagnostic potential of various antigenic proteins of Fasciola gigantica species isolated from sheep of North West Himalayan Region

Immunolocalization and immunodetection of the excretory/secretory (ES) antigens of Fasciola gigantica

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