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

  • Kamel, Hanan H. (Department of Parasitology, Faculty of Medicine, Ain Shams University) ;
  • Saad, Ghada A. (Department of Parasitology, Faculty of Medicine, Ain Shams University) ;
  • Sarhan, Rania M. (Department of Parasitology, Faculty of Medicine, Ain Shams University)
  • Received : 2012.10.29
  • Accepted : 2013.01.30
  • Published : 2013.04.30

Abstract

The purpose of the present study was to evaluate the potential role of the 27-Kilodalton (KDa) antigen versus Fasciola gigantica adult worm regurge antigens in a DOT-Blot assay and to assess this assay as a practical tool for diagnosis fascioliasis in Egyptian patients. Fasciola gigantica antigen of an approximate molecular mass 27- (KDa) was obtained from adult worms by a simple elution SDS-PAGE. A Dot-Blot was developed comparatively to adult worm regurge antigens for the detection of specific antibodies from patients infected with F. gigantica in Egypt. Control sera were obtained from patients with other parasitic infections and healthy volunteers to assess the test and compare between the antigens. The sensitivity, specificity, positive and negative predictive values of Dot-Blot using the adult worm regurge were 80%, 90%, 94.1%, and 69.2% respectively, while those using 27-KDa were 100% which confirms the diagnostic potential of this antigen. All patients infected with Fasciola were positive, with cross reactivity reported with Schistosoma mansoni serum samples. This 27-KDa Dot-Blot assay showed to be a promising test which can be used for serodiagnosis of fascioliasis in Egyptian patients especially, those presenting with hepatic disease. It is specific, sensitive and easy to perform method for the rapid diagnosis particularly when more complex laboratory tests are unavailable.

Keywords

References

  1. Schweizer G, Braun U, Deplazes P, Torgerson PR. Estimating the financial losses due to bovine fasciolosis in Switzerland. Vet Rec 2005; 157: 188-193.
  2. Mungube EO, Bauni SM, Tenhagen BA, Wamae LW, Nginyi JM, Mugambi JM. The prevalence and economic significance of Fasciola gigantica and Stilesia hepatica in slaughtered animals in the semi-arid coastal Kenya. Trop Anim Health Prod 2006; 38, 475-483. https://doi.org/10.1007/s11250-006-4394-4
  3. Mas-Coma MS, Esteban JG, Bargues MD. Epidemiology of human fascioliasis: a review and proposed new classification. Bull World Health Organ 1999; 77: 340-346.
  4. World Health Organisation. Control of foodborne trematode infections. Report of a WHO Study Group. World Health Organ Tech Rep Ser 1995; 849: 1-157.
  5. Andrews SJ. The life cycle of Fasciola hepatica. In Dalton JP ed, Fasciolosis. Walling Ford, UK. CABI Publishing. 1999, p 1-20.
  6. Mas-Coma S, Bargues MD, Valero MA. Fascioliasis and other plant-borne trematode zoonoses. Int J Parasitol 2005; 35: 1255-1278. https://doi.org/10.1016/j.ijpara.2005.07.010
  7. Maleewong W, Wongkham C, Intapan PM, Pipitgool V. Fasciola gigantica-specific antigens: purification by a continuous-elution method and its evaluation for the diagnosis of human fascioliasis. Am J Trop Med Hyg 1999; 61: 648-651.
  8. De Almeida MA, Ferreira MB, Planchart S, Terashima A, Maco V, Marcos L, Gotuzzo E, Sanchez E, Naquira C, Scorza JV, Incani RN. Preliminary antigenic characterisation of an adult worm vomit preparation of Fasciola hepatica by infected human sera. Rev Inst Med Trop Sao Paulo 2007; 49: 31-35.
  9. Planchart S, Incani RN, Ballen D, Matos C, Cesari I. Preliminary biochemical characterization of a Schistosoma mansoni adult worm vomit (AWV) preparation and immunological specificity of a mouse anti-AWV monoclonal antibody. In International symposium on schistosomiasis, 9th. Salvador. 2003.
  10. Jeno P, Horst M. Electroelution of Proteins from Polyacrylamide Gels. In Walker JM ed, The Protein Protocols Handbook. 2nd ed. Totowa, New Jersey, USA. Humana Press. 2002, p 207-214.
  11. Lammeli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685. https://doi.org/10.1038/227680a0
  12. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein- dye binding. Anal Biochem 1976; 72: 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
  13. Wright SF, Morton JB. Detection of vesicular-arbuscular mycorrhizal fungus colonization of roots by using a dot-immunoblot assay. Appl Environ Microbiol 1989; 55: 761-763.
  14. Bayu B, Asnake S, Woretaw A, Ali J, Gedefaw M, Fente T, Getachew A, Tsegaye S, Dagne T, Yitayew G. Cases of human fascioliasis in North-West Ethiopia. Ethiop J Health Dev 2005; 19: 237-240.
  15. Marcos L, Romani L, Florencio L, Terashima A, Canales M, Nestares J, Huayanay L, Gotuzzo E. Hyperendemic and mesoendemic zones of Fasciola infection surrounding urban Lima: an emerging disease? Rev Gastroenterol Peru 2007; 27: 31-36.
  16. Marcos LA, Tagle M, Terashima A, Bussalleu A, Ramirez C, Carrasco C, Valdez L, Huerta-Mercado J, Freedman DO, Vinetz JM, Gotuzzo E. Natural history, clinicoradiologic correlates, and response to triclabendazole in acute massive fascioliasis. Am J Trop Med Hyg 2008; 78: 222-227.
  17. Mas-Coma S, Bargues MD, Esteban JG. Human fasciolosis. In Dalton JP ed, Fasciolosis. Walling Ford, UK. CABI Publishing. 1999, p 411-433.
  18. Rahimi MT, Ashrafi K, Koosha S, Abdi J, Rokni MB. Evaluation of Fast-ELISA versus standard-ELISA to diagnose human fasciolosis. Arch Iran Med 2011; 14: 18-21.
  19. Espino AM, Diaz A, Perez A, Finlay CM. Dynamics of antigenemia and coproantigens during a human Fasciola hepatica outbreak. J Clin Microbiol 1998; 36: 2723-2726.
  20. Almazan C, Avila G, Quiroz H, Ibarra F, Ochoa P. Effect of parasite burden on the detection of Fasciola hepatica antigens in sera and feces of experimentally infected sheep. Vet Parasitol 2001; 97: 101-112. https://doi.org/10.1016/S0304-4017(01)00376-4
  21. Dixit AK, Yadav SC, Sharma RL. 28 kDa Fasciola gigantica cysteine proteinase in the diagnosis of prepatent ovine fasciolosis. Vet Parasitol 2002; 109: 233-247. https://doi.org/10.1016/S0304-4017(02)00202-9
  22. Maleewong W, Intapan PM, Wongkham C, Tomanakan K, Daenseekaew W, Sukeepaisarnjaroen W. Comparison of adult somatic and excretory-secretory antigens in enzyme-linked immunosorbent assay for serodiagnosis of human infection with Fasciola gigantica. Southeast Asian J Trop Med Public Health 1996; 27: 566-569.
  23. Dalimi A, Hadighi R, Madani R. Partially purified fraction (PPF) antigen from adult Fasciola gigantica for the serodiagnosis of human fascioliasis using Dot-ELISA technique. Ann Saudi Med 2004; 24: 18-20.
  24. Silva E, Castro A, Lopes A, Rodrigues A, Dias C, Conceicao A, Alonso J, Correia da Costa JM, Bastos M, Parra F, Moradas-Ferreira P, Silva M. A recombinant antigen recognized by Fasciola hepatica-infected hosts. J Parasitol 2004; 90: 746-751. https://doi.org/10.1645/GE-136R
  25. Yamasaki H, Aoki T, Oya H. A cysteine proteinase from the liver fluke Fasciola spp.: purification, characterization, localization and application to immunodiagnosis. Jpn J Parasitol 1989; 38: 373-384.
  26. Cordova M, Herrera P, Nopo L, Bellatin J, Naquira C, Guerra H, Espinoza JR. Fasciola hepatica cysteine proteinases: immunodominant antigens in human fascioliasis. Am J Trop Med Hyg 1997; 57: 660-666.
  27. Sampaio-Silva ML, Da Costa JM, Da Costa AM, Pires MA, Lopes SA, Castro AM, Monjour L. Antigenic components of excretorysecretory products of adult Fasciola hepatica recognized in human infections. Am J Trop Med Hyg 1996; 54: 146-148.
  28. Sripa B, Kaewkes S, Sithithaworn P, Mairiang E, Laha T, Smout M, Pairojkul C, Bhudhisawasdi V, Tesana S, Thinkamrop B, Bethony JM, Loukas A, Brindley PJ. Liver fluke induces cholangiocarcinoma. PLoS Med 2007; 4: e201. https://doi.org/10.1371/journal.pmed.0040201
  29. Nguyen TG, Le TH, De NV, Doan TT, Dao TH, Vercruysse J, Dorny P. Assessment of a 27-kDa antigen in enzyme-linked immunosorbent assay for the diagnosis of fasciolosis in Vietnamese patients. Trop Med Int Health 2010; 15: 462-467.
  30. Farghaly AM, Nada SMM, Emam WA, Mattar MA, Mohamed SMA, Sharaf EM, El Gamal RLE. Role of fast-ELISA and western blot in diagnosis of human fascioliasis using crude adult worm and excretory/secretory Fasciola antigens. Parasitologists United Journal (PUJ) 2009; 2: 55- 65.
  31. Abdel-Rahman EH, Abdel-Megeed KN, Abuel-Ezz NM. Cross-reaction: a common trait among helminthes. J Egypt Soc Parasitol 2003; 33: 457-471.

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