Development and Application of a Fecal Antigen Diagnostic Sandwich ELISA for Estimating Prevalence of Fasciola gigantica in Cattle in Central Java, Indonesia

Indonesian Research Institute for Veterinary Science, Bogor, West Java, Indonesia.
Journal of Parasitology (Impact Factor: 1.23). 09/2008; 95(2):450-5. DOI: 10.1645/GE-1672.1
Source: PubMed


The purpose of this study was to compare the sensitivity and specificity of an ELISA test to detect Fasciola gigantica antigens (coproantigens) in bovine feces, with fecal egg counting and an ELISA for detecting anti-F. gigantica antibodies in serum. Monoclonal antibodies to cathepsin L were generated and used to capture this antigen in feces of infected cattle. Blood, feces, and livers were collected from 150 cattle at an abattoir in Jakarta, Indonesia, for anti-Fasciola antibodies, coproantigen detection, and F. gigantica egg and worm counts. Fluke recovery varied from 1 to 426 per host, with a mean of 32 flukes. The results showed that the sensitivity and specificity of coproantigen detecting ELISA (95 and 91%, respectively) was better than the anti-F. gigantica antibody ELISA (91 and 88%, respectively) and to fecal egg counting (87 and 100%, respectively). The coproantigen ELISA was able to detect 100% of the cattle with >15 flukes. A survey of 305 cattle in central Java over a 10-mo period validated this test in the field, demonstrating a high prevalence of fascioliasis and establishing the test as a useful diagnostic method to determine patent F. gigantica infections in cattle.

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Available from: Ruby HP Law, Dec 27, 2013
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    • "An alternative to antibody detection is the direct measurement of F. gigantica antigens that are shed into the serum or faeces of infected animals or humans (Valero et al., 2009). A monoclonal antibody has been produced that can detect F. gigantica antigen in the serum and faeces of infected humans (Demerdash et al., 2011) and cattle (Estuningsih et al., 2009). Antibody detection assays are most commonly used due to the relative simplicity of the assays, and early seroconversion (usually 1 – 2 weeks), high sensitivity and easy usage (Jalali et al., 2012). "
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    ABSTRACT: A serological and coprological survey of fasciolosis was conducted in bovine hosts from the Sargodha district, Pakistan using excretory–secretory (ES) antigens of Fasciola gigantica from cattle and buffaloes. Livers, faecal and blood samples of 146 cattle and 184 buffaloes were collected from slaughterhouses and examined for the presence of any Fasciola in bile ducts and ova in faeces. Serum was separated. ES antigens were prepared by incubating adult Fasciola in phosphate-buffered saline for 6–8 h and then filtering using a 0.22-μm syringe filter. Checkerboard titration was performed and optimum concentrations of antigen and serum were determined. Sero-prevalence was found to be 50.00 and 38.35% in buffalo and cattle, respectively. Using liver examination as the gold standard, enzyme-linked immunosorbent assay (ELISA) sensitivity was found to be 100% in both buffalo and cattle as compared with that of coprological examination in buffalo (61.79%) and cattle (54.54%). This indigenous ELISA was also highly specific, with values of 96.84 and 98.90% in buffalo and cattle, respectively. Positive predictive values were calculated as 96.74 and 98.21% in buffalo and cattle, respectively, while negative predictive values were 100%. For the validation of indigenous ELISA in field surveys, faecal and blood samples were collected from six sub-districts (tehsils) in the district of Sargodha. Sera were screened for the presence of anti-fasciola antibodies using both the indigenous and commercial ELISA kits. While both kits were equally sensitive, the indigenous ELISA was found to be more specific. The highest prevalence of fasciolosis was found in December, as ascertained using both serological and coprological examination. Significant differences were found in prevalences of fasciolosis in different sub-districts and age groups, together with feeding and watering systems.
    Full-text · Article · Aug 2015 · Journal of Helminthology
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    • "causes major economic losses to Australian (A$60–90 million p.a.) and global (>US$3 billion p.a.) livestock production with over 600 million animals infected (Spithill et al., 1999a;Mas-Coma et al., 2005;Piedrafita et al., 2010). In the tropics, fasciolosis is recognised as a major helminth infection of cattle (Fabiyi, 1987) with prevalence rates ranging between 25–100% in Africa, the Middle East, Asia and southeastern Asia (reviewed inSchillhorn van Veen, 1980;Spithill et al., 1999a;Estuningsih et al., 2009). In developed countries, the incidence of F. hepatica can be as high as 77% (Wilson et al., 1982;Dargie, 1987) with prevalences up to 95% recently observed in dairy cattle in Australia (Spithill et al., unpublished observations). "
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    ABSTRACT: The development of a vaccine for Fasciola spp. in livestock is a challenge and would be advanced by harnessing our knowledge of acquired immune mechanisms expressed by resistant livestock against fluke infection. Antibody-dependent cell-mediated cytotoxicity directed to the surface tegument of juvenile/immature flukes is a host immune effector mechanism, suggesting that antigens on the surface of young flukes may represent prime candidates for a fluke vaccine. A Type 1 immune response shortly after fluke infection is associated with resistance to infection in resistant sheep, indicating that vaccine formulations should attempt to induce Type 1 responses to enhance vaccine efficacy. In cattle or sheep, an optimal fluke vaccine would need to reduce mean fluke burdens in a herd below the threshold of 30 - 54 flukes to ensure sustainable production benefits. Fluke infection intensity data suggest that vaccine efficacy of approximately 80% is required to reduce fluke burdens below this threshold in most countries. With the increased global prevalence of triclabendazole-resistant F. hepatica, it may be commercially feasible in the short term to introduce a fluke vaccine of reasonable efficacy that will provide economic benefits for producers in regions where chemical control of new drug-resistant fluke infections is not viable. Commercial partnerships will be needed to fast-track new candidate vaccines using acceptable adjuvants in relevant production animals, obviating the need to evaluate vaccine antigens in rodent models.
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    • "Several coproantigen capture ELISAs have been developed to diagnose trematode infections in humans and animals [20] [21] [22] [23] [24] [25] [26] [27] [28] [29]. The advantage of these ELISAs is that the parasite antigens can be detected in the feces before eggs are present, which facilitates the diagnosis of early or latent infection. "
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    ABSTRACT: The present study investigated the diagnostic value of an ELISA for the detection of Clonorchis sinensis antigen in the feces of experimentally infected rats. A mouse polyclonal IgG antibody against adult C. sinensis crude antigen (CsAg) was used to capture the C. sinensis coproantigen. The detection limit for pure CsAg was 20 ng/ml in sample buffer and 40 ng/ml in uninfected fecal extract. The test was evaluated using a follow-up of five groups of rats experimentally infected with 100, 50, 10, 5 and 1 metacercariae of C. sinensis and an uninfected control group. Coproantigen was detected in all infected groups of rats from 2 weeks of infection, whereas fecal eggs were not observed until 3 weeks of infection. As the infection period progressed, the fecal CsAg concentration increased in all groups of infected rats, even those infected with a single metacercaria. The fecal CsAg concentration was correlated positively with fecal egg counts and worm burden. This coproantigen capture ELISA is highly sensitive for the detection of CsAg in rat feces, and with further development, should be useful for mass screening of human subjects in clonorchiasis-endemic areas.
    Full-text · Article · Aug 2011 · Parasitology International
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