Use of the PCR in a Combined Methodological Approach for the Study of Human Fascioliasis in an Endemic Area



Fascioliasis is a worldwide distributed trematodiasis considered a neglected disease. Diagnosis in humans has been traditionally based on parasitological and immunological techniques. Recently we reported the use of the PCR in stool samples for the individual diagnosis. The purpose of this study was to evaluate human fascioliasis by a combination of diagnostic methods in an area where the disease is highly endemic in animals.


We studied all the inhabitants (N = 240) of Tatón village, Argentina, by Fasciola hepatica rproCL1-ELISA. Among them, we continued the study with 13 cases that had at least two positive serological tests, who performed a questionnaire, physical examination, abdominal ultrasonography, and collection of blood and faeces. Blood/serum samples were used for Fh rproCL1-ELISA and liver function tests. Faeces were used for parasitological analysis and PCR of a repetitive fragment of Fasciola sp.


Among the 13 patients, 9 presented symptoms of biliary colic. All patients repeated positive serology. F. hepatica eggs were not detected. PCR was positive in 11 cases.


This is the first report employing an approach based on the combination of methods for the evaluation of human fascioliasis in an endemic area, which includes molecular tools with a high value in detecting low infections.

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  1. 1.

    Arjona R, Riancho J, Aguado J, Salesa R, Gonzalez-Macias J (1995) Fascioliasis in developed countries: a review of classic and aberrant forms of the disease. Medicine (Baltimore) 74:13–23.

    CAS  Article  Google Scholar 

  2. 2.

    Binkley CE, Sinniah B (1997) Fascioliasis. In: Connor DH, Chandler FW, Schwartz DA, Manz HJ, Lack EE (eds) Pathology of infectious diseases. Appleton and Lange, Stamford, pp 1419–1425

    Google Scholar 

  3. 3.

    Bryan RT, Michelson MK (1995) Parasitic infections of the liver and biliary tree. In: Surawicz C, Owen RL (eds) Gastrointestinal and hepatic infections. W.B. Saunders, Philadelphia, pp 405–454

    Google Scholar 

  4. 4.

    Cabada MM, Malaga JL, Castellanos-Gonzalez A, Bagwell KA, Naeger PA, Rogers HK, Maharsi S, Mbaka M, White AC Jr (2017) Recombinase polymerase amplification compared to Real-Time polymerase chain reaction test for the detection of Fasciola hepatica in human stool. Am J Trop Med Hyg 96:341–346.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  5. 5.

    Carnevale S, Cabrera MG, Cucher MA, di Risio CA, Malandrini JB, Kamenetzky L, Alazraqui M, Etchart CB, Pantano ML, Velásquez JN (2013) Direct, immunological and molecular techniques for a fasciolosis survey in a rural area of San Luis, Argentina. J Parasit Dis 37:251–259.

    Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Carnevale S, Malandrini JB, Pantano ML, Sawicki M, Soria CC, Kuo LH, Kamenetzky L, Astudillo OG, Velásquez JN (2016) Fasciola hepatica infection in humans: overcoming problems for the diagnosis. Acta Parasitol 61:776–783.

    Article  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Carnevale S, Rodríguez MI, Guarnera EA, Carmona C, Tanos T, Angel S (2001) Immunodiagnosis of fasciolosis using recombinant procathepsin L cystein proteinase. Diagn Microbiol Infect Dis 41:43–49.

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Carnevale S, Rodríguez MI, Santillan G, Labbé JH, Cabrera MG, Bellegarde EJ, Velásquez JN, Trgovcic JE, Guarnera EA (2001) Immunodiagnosis of human fascioliasis by enzyme-linked immunosorbent assay (ELISA) and Micro-ELISA. Clin Diagn Lab Immunol 8:174–177.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. 9.

    Chen MG, Mott KE (1990) Progress in assessment of morbidity due to Fasciola hepatica infection: a review of recent literature. Trop Dis Bull 87:R1–R38

    Google Scholar 

  10. 10.

    Espino Hernandez AM, Seuret N, Morlote C, Dumenigo Ripoll B (1991) Antígenos de Fasciola hepatica. Su utilidad en el diagnóstico de la fascioliasis humana. Rev Cubana Med Trop 43:151–155

    CAS  PubMed  Google Scholar 

  11. 11.

    Espinoza JR, Maco V, Marcos L, Saez S, Neyra V, Terashima A, Samalvides F, Gotuzzo E, Chavarry E, Huaman MC, Bargues MD, Valero MA, Mas-Coma S (2007) Evaluation of Fas2-ELISA for the serological detection of Fasciola hepatica infection in humans. Am J Trop Med Hyg 76:977–982

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Espinoza JR, Timoteo O, Herrera-Velit P (2005) Fas2-ELISA in the detection of human infection by Fasciola hepatica. J Helminthol 79:235–240.

    CAS  Article  PubMed  Google Scholar 

  13. 13.

    Hillyer GV, Soler de Galanes M, Rodriguez-Perez J, Bjorland J, De Lagrava MS, Ramirez Guzman S, Bryan RT (1992) Use of the Falcon assayTM screening test-enzyme linked immunosorbent assay (FAST-ELISA) and the enzyme-linked immunoelectrotransfer blot (EITB) to determine the prevalence of human fascioliasis in the Bolivian Altiplano. Am J Trop Med Hyg 46:603–609.

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Knobloch J (1985) Human fascioliasis in Cajamarca/Peru: humoral antibody response and antigenaemia. Trop Med Parasitol 36:91–93

    CAS  PubMed  Google Scholar 

  15. 15.

    L’Ollivier C, Eldin C, Lambourg E, Brouqui P, Lagier JC (2020) Case report: First molecular diagnosis of liver abscesses due to Fasciola hepatica acute infection imported from Vietnam. Am J Trop Med Hyg 102:106–109.

    Article  PubMed  Google Scholar 

  16. 16.

    Mas-Coma S, Bargues MD, Valero MA (2005) Fascioliasis and other plant-borne trematode zoonoses. Int J Parasitol 35:1255–1278.

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Mas-Coma S, Bargues MD, Valero MA (2014) Diagnosis of human fascioliasis by stool and blood techniques: update for the present global scenario. Parasitology 141:1918–1946.

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Mas-Coma S, Funatsu IR, Bargues MD (2001) Fasciola hepatica and lymnaeid snails occurring at very high altitude in South America. Parasitology 123:S115–S127.

    Article  PubMed  Google Scholar 

  19. 19.

    Mas-Coma S, Valero MA, Bargues MD (2019) Fascioliasis. In: Toledo R, Fried B (eds) Digenetic trematodes. Advances in experimental medicine and biology. Springer, Cham, pp 71–103.

    Google Scholar 

  20. 20.

    O’Neill SM, Parkinson M, Dowd AJ, Strauss W, Angles R, Dalton JP (1999) Short report: immunodiagnosis of human fascioliasis using recombinant Fasciola hepatica cathepsin L1 cysteine proteinase. Am J Trop Med Hyg 60:749–751.

    Article  PubMed  Google Scholar 

  21. 21.

    O’Neill SM, Parkinson M, Strauss W, Angles R, Dalton JP (1998) Immunodiagnosis of Fasciola hepatica infection (fascioliasis) in a human population in the Bolivian Altiplano using purified cathepsin L cysteine proteinase. Am J Trop Med Hyg 58:417–423.

    Article  PubMed  Google Scholar 

  22. 22.

    Quiroga A (1986) Excursiones por Pomán y Tinogasta. Valles de Abaucán (provincia de Catamarca). Bol del Inst Geogr Argent 17:499–526

    Google Scholar 

  23. 23.

    Raymundo LA, Maco Flores V, Terashima A, Samalvides F, Miranda E, Tantalean M, Espinoza JR, Gotuzzo E (2004) Hiperendemicidad de Fasciolosis humana en el Valle del Mantaro, Perú: factores de riesgo de la infeccion por Fasciola hepatica. Rev Gastroenterol Peru 24:158–164

    PubMed  Google Scholar 

  24. 24.

    Trueba G, Guerrero T, Fornasini M, Casariego I, Zapata S, Ontaneda S, Vasco L (2000) Detection of Fasciola hepatica infection in a community located in the Ecuadorian Andes. Am J Trop Med Hyg 62:518.

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Valero MA, Periago MV, Pérez-Crespo I, Angles R, Villegas F, Aguirre C, Strauss W, Espinoza JR, Herrera P, Terashima A, Tamayo H, Engels D, Gabrielli AF, Mas-Coma S (2012) Field evaluation of a coproantigen detection test for fascioliasis diagnosis and surveillance in human hyperendemic areas of Andean countries. PLoS Negl Trop Dis 6:e1812.

    Article  PubMed  PubMed Central  Google Scholar 

  26. 26.

    Zumaquero-Ríos JL, Sarracent-Pérez J, Rojas-García R, Rojas-Rivero L, Martínez-Tovilla Y, Valero MA, Mas-Coma S (2013) Fascioliasis and intestinal parasitoses affecting schoolchildren in Atlixco, Puebla State, Mexico: epidemiology and treatment with nitazoxanide. PLoS Negl Trop Dis 7:e2553.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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Correspondence to Jorge Néstor Velásquez.

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Carnevale, S., Malandrini, J.B., Pantano, M.L. et al. Use of the PCR in a Combined Methodological Approach for the Study of Human Fascioliasis in an Endemic Area. Acta Parasit. (2020).

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  • Human fascioliasis
  • Polymerase chain reaction
  • Enzyme-linked immunosorbent assay
  • Endemic areas