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.
    Journal of Helminthology 08/2015; -1:1-9. DOI:10.1017/S0022149X15000711 · 1.42 Impact Factor
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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.
    Parasitology International 08/2011; 61(1):203-7. DOI:10.1016/j.parint.2011.08.006 · 1.86 Impact Factor
    • "Current tests for immunodiagnosis of human Fasciola infection include ELISAs with ES antigens or purified/recombinant proteins in serum (Dixit et al., 2008; Espinoza et al., 2005; O'Neill et al., 1999). Detection of Fasciola antigens in F. gigantica-infected mouse serum (Anuracpreeda et al., 2009), and coproantigens in faeces from infected cattle (Estuningsih et al., 2009) and humans (Ubeira et al., 2009) has been reported. The latter using MM3-COPRO method with a new preservative/diluent, CoproGuard, for preservation of Fasciola coproantigens is promising with 100% sensitivity and 100% specificity. "
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    ABSTRACT: The food-borne trematodiases are an important group of neglected tropical diseases (NTDs). Over 40 million people are infected with food-borne trematodes and 750 million (>10% of the world's population) are at risk of these NTDs. Here, we review the life cycles, epidemiology, clinical manifestations, pathology and pathogenesis, diagnosis, treatment, and prevention and control of the major food-borne trematodiases in Southeast Asia. We focus particularly on opisthorchiasis caused by Opisthorchis viverrini and clonorchiasis caused by Clonorchis sinensis, which people contract by ingestion of metacercariae in flesh of raw or undercooked freshwater fishes, on fascioliasis caused by Fasciola species, where infection arises from ingestion of metacercariae on water plants such as watercress, and on Paragonimus species, the lung flukes, which use freshwater crabs and other crustaceans as intermediate hosts. We also include information on the intestinal flukes Fasciolopsis buski, the echinostomes and the so-called 'minute intestinal flukes' of the family Heterophyidae. Ecological information, placing emphasis on reservoir hosts, intermediate snail hosts and secondary hosts where applicable, is also reviewed and research needs are highlighted.
    Advances in Parasitology 01/2010; 72:305-50. · 6.23 Impact Factor
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