Characterization of specific and cross-reacting antigens of Fasciola gigantica by immunoblotting

Yokananth, S.; Ghosh, Srikant; Gupta, Sandeep; Suresh, Murugan; Saravanan, D.

doi:10.1007/s00436-005-1371-1

Cited by 17 publications

(15 citation statements)

References 27 publications

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“…Allam et al (2002) and Gonence and Sarimehmetoglu (2004) in two different studies reported fewer protein bands in SDS-PAGE pattern of Fasciola in comparison to our study. Study of Yokananth et al (2005), about electrophoretic separation of F. gigantica somatic antigen, showed 20 protein bands (ranging from 14 to 156 kDa) (Yokananth et al 2005). Eight protein bands reported in their study are common with those that we are seeing in the current study.…”

Section: Discussionsupporting

confidence: 73%

“…It has been shown that sera of cattle, sheep and buffalo, infected with F. gigantica, reacted respectively with 5, 8 and 11 protein bands of F. gigantica somatic antigen (Yokananth et al 2005). In the present study rabbit polyclonal antibodies raised against F. gigantica somatic and ES antigen respectively reacted with 19 and 21 protein bands of F. gigantica somatic antigen.…”

Section: Discussionsupporting

confidence: 57%

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Detection of Fasciola hepatica and Fasciola gigantica common and uncommon antigens, using rabbit hyper immune serum raised against their excretory–secretory and somatic antigens

Khabisi

Sarkari

2016

J Parasit Dis

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Fasciolosis is an important neglected helminth disease caused by two liver flukes, Fasciola hepatica and Fasciola gigantica. The two species of Fasciola are usually different in their morphological and molecular features. They have also common and uncommon antigens in both their somatic and excretory secretory metabolites. In this study, we compared somatic and excretory-secretory (ES) antigens of F. hepatica and F. gigantica, by using rabbit hyper immune serum raised against these antigens. Adult worms were collected from bile ducts of infected animals and species of the fluke was confirmed by RFLP-PCR. ES and somatic antigens of both species were prepared. Rabbits were subcutaneously immunized with either ES or somatic antigens to produce antibodies against these antigens. SDS-PAGE pattern of F. hepatica and F. gigantica somatic antigens was similar and both of them revealed 30 protein bands, ranging from 18 to 180 kDa. In contrast, SDS-PAGE pattern of ES antigen of the two species was different. While protein bands with molecular weight of 18, 27, 29, 48, and 62 kDa were common in both species, bands of 19, 45, 55 and 58 kDa were only noticed in F. hepatica ES antigen. Rabbit polyclonal antibodies, raised against F. hepatica and F. gigantica ES antigen, reacted with main five protein bands, 25, 27, 29, 62 and 67 kDa and polyclonal antibodies raised against somatic antigens of both species reacted with three protein bands, 25, 27 and 72 kDa. Thus, the 25, 27 and 29 kDa protein bands may serve as immunodominant antigens, which might be considered for serodiagnosis of fasciolosis. Moreover, bands of 62 and 67 kDa in ES antigen and 72 kDa in somatic antigens of both species were immunodominant and might be suitable candidate for development of serological assays for diagnosis of fasciolosis.

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Section: Discussionsupporting

confidence: 73%

Section: Discussionsupporting

confidence: 57%

Detection of Fasciola hepatica and Fasciola gigantica common and uncommon antigens, using rabbit hyper immune serum raised against their excretory–secretory and somatic antigens

Khabisi

Sarkari

2016

J Parasit Dis

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“…The affinity purified antigens for Fasciola spp. have shown high sensitivity and specificity (Fagbemi and Guobadia 1995;Intapan et al 2003;Ghosh et al 2005;Yokananth et al 2005). Hence, on similar lines affinity chromatography was employed in the present study for the purification of specific immunodominant polypeptides against P. epiclitum.…”

Section: Resultsmentioning

confidence: 90%

“…However, till date no specific diagnostic polypeptide against P. epiclitum has been identified and purified. Affinity chromatography has been shown to be a very effective tool for isolation of candidate diagnostic and vaccine molecules (Ghosh et al 1999;Sharma et al 2001;Singh and Ghosh 2003;Yokananth et al 2005). The affinity purified antigens for Fasciola spp.…”

Section: Resultsmentioning

confidence: 99%

Immunoaffinity chromatographic analysis for purification of specific diagnostic antigens of Paramphistomum epiclitum

et al. 2010

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Polypeptide profile of somatic antigen of Paramphistomum epiclitum (PSAg) and Gastrothylax crumenifer (GSAg) was studied by SDS-PAGE. PSAg and GSAg showed 14 and 19 polypeptides in the range of 14.9-95.5 and 13.7-129.6 kDa with six common polypeptides of mol wt 16.8, 21.8, 23.7, 35.5, 43.4 and 70.8 kDa. P. epiclitum experimentally infected sheep sera were used for identification of specific immuno-dominant peptide in the range of 37-40 kDa against P. epiclitum by western blotting. Hyperimmune sera (HIS) was raised in rabbit against the identified polypeptide, IgG was separated from HIS and an immunoaffinity column was constructed with a binding percentage of 83.74 of IgG with CNBr activated Sepharose 4B. Purification of somatic antigen (PSAg) was done with immunoaffinity chromatography and 37-40 kDa protein antigen was isolated in pure form with recovery percentage of 2.97%. This purified fraction of somatic antigen can be used as a candidate antigen for development of serological assay for early diagnosis of paramphistomosis among livestock.

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“…This study reports an assessment of two recombinant subunits of the antigen B, B1t and B2t, and the use of two new recombinant antigens, E14t and C317, in an ELISA for the detection of specific antibodies in UHD patients. For this study, we selected a well-characterized and extensive panel of sera that included sera from patients with UHD (n ϭ 102) confirmed by surgery; from patients with different parasitic diseases (n ϭ 68), especially those related to E. granulosus at the antigen composition level (alveolar hydatid disease, cysticercosis, teniosis, schistosomosis, fasciolosis, and nematodes) (7,18,30); and healthy donors (n ϭ 95). This latter group of individuals was chosen from an area free of E. granulosus, because healthy donors from areas in which E. granulosus is endemic could give rise to false-positive results from individuals who have been in contact with the parasite without developing the disease (32).…”

Section: Vol 15 2008 Serodiagnosis and Follow-up Of Hydatid Diseasementioning

confidence: 99%

Usefulness of Four Different Echinococcus granulosus Recombinant Antigens for Serodiagnosis of Unilocular Hydatid Disease (UHD) and Postsurgical Follow-Up of Patients Treated for UHD

Hernández-González

Muro

Barrera

et al. 2008

Clin Vaccine Immunol

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Four different recombinant antigens derived from Echinococcus granulosus, designated B1t, B2t, E14t, and C317, were tested with enzyme-linked immunosorbent assays (ELISAs) for the detection of specific immunoglobulin G (IgG) in patients with unilocular hydatid disease (UHD). The results were compared to those obtained with hydatid fluid and were subjected to receiver operator characteristic analysis. The diagnostic performance of the above-listed proteins was defined with respect to their specificity, sensitivity, and predictive values (PV); the influence of cyst location; and usefulness in the follow-up of surgical treatment for UHD and in the determination of whether or not patients have been surgically cured of UHD. The best diagnostic results were obtained with the anti-B2t IgG ELISA, with 91.2% sensitivity, 93% specificity, and high positive and negative PV (89.4 and 94.2, respectively). In addition, this diagnostic tool proved to be useful for the follow-up of surgically treated UHD patients. The anti-B2t IgG ELISA may find an application in the serodiagnosis of UHD in clinical laboratories.Human unilocular hydatid disease (UHD) is caused by the larvae of the tapeworm Echinococcus granulosus. The diagnosis of UHD usually is approached by applying an imaging technique. In 2003, the World Health Organization Informal Working Group on Echinococcosis (28) proposed the use of ultrasonography (US) as the imaging technique of choice in order to promote uniform standards of UHD diagnosis and follow-up. It also was pointed out that US images suspected of showing UHD should be examined by alternative diagnostic methods, such as serological techniques (28). In addition, for pulmonary UHD, US is unhelpful in most cases, unless the cysts are close to the pleural surface (12). Currently, the preferred UHD immunodiagnostic techniques used in clinical practice detect serum immunoglobulin G (IgG) against crude parasite extracts (mainly hydatid fluid [HF]), most commonly by hemagglutination and enzyme-linked immunosorbent assay (ELISA) (26). However, HF cannot be standardized and gives rise to relatively frequent false-positive and false-negative results (3, 15). Moreover, HF is not useful for the follow-up of UHD, since the levels of specific (IgG) antibodies against HF remain high over long periods of time after curing (19). Many authors have attempted to find alternative antigens to improve UHD serodiagnosis and follow-up. Thus, purified, recombinant antigens and synthetic peptides, mainly derived from two parasite molecules (antigen B [AgB] and antigen 5 [Ag5]), have been applied, with various levels of success. Specifically, antigens derived from AgB have been shown to be good candidates for UHD serodiagnosis (3). AgB is formed by several subunits (6), two of which (AgB1 and AgB2) have been produced as recombinant molecules. Both AgB1 and AgB2 have been tested for the detection of specific antibodies, although authors disagree about the diagnostic performance of these two subunits (11,20,27). Thus, the standardization ...

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Characterization of specific and cross-reacting antigens of Fasciola gigantica by immunoblotting

Cited by 17 publications

References 27 publications

Detection of Fasciola hepatica and Fasciola gigantica common and uncommon antigens, using rabbit hyper immune serum raised against their excretory–secretory and somatic antigens

Detection of Fasciola hepatica and Fasciola gigantica common and uncommon antigens, using rabbit hyper immune serum raised against their excretory–secretory and somatic antigens

Immunoaffinity chromatographic analysis for purification of specific diagnostic antigens of Paramphistomum epiclitum

Usefulness of Four Different Echinococcus granulosus Recombinant Antigens for Serodiagnosis of Unilocular Hydatid Disease (UHD) and Postsurgical Follow-Up of Patients Treated for UHD

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