Skip to main content

Advertisement

SpringerLink
Log in

Serological diagnosis of echinococcosis: the diagnostic potential of native antigens

  • Clinical and Epidemiological Study
  • Published:
Infection Aims and scope Submit manuscript

Abstract

Purpose

Human alveolar (AE) and cystic echinococcosis (CE) caused by the metacestode stages of Echinococcus multilocularis and E. granulosus, respectively, lack pathognomonic clinical signs. Diagnosis therefore relies on the results of imaging and serological studies. The primary goal of this study was to evaluate the efficacy of several easy-to-produce crude or partially purified E. granulosus and E. multilocularis metacestode-derived antigens as tools for the serological diagnosis and differential diagnosis of patients suspicious for AE or CE.

Methods

The sera of 51 treatment-naïve AE and 32 CE patients, 98 Swiss blood donors and 38 patients who were initially suspicious for echinococcosis but suffering from various other liver diseases (e.g., liver neoplasia, etc.) were analysed.

Results

According to the results of enzyme-linked immunosorbent assays (ELISA), metacestode-derived antigens of E. granulosus had sensitivities varying from 81 to 97% and >99.9% for the diagnosis of CE and AE, respectively. Antigens derived from E. multilocularis metacestodes had sensitivities ranging from 84 to 91% and >99.9% for the diagnosis of CE and AE, respectively. Specificities ranged from 92 to >99.9%. Post-test probabilities for the differential diagnosis of AE from liver neoplasias, CE from cystic liver lesions, and screening for AE in Switzerland were around 95, 86 and 2.2%, respectively. Cross-reactions with antibodies in sera of patients with other parasitic affections (fasciolosis, schistosomosis, amebosis, cysticercosis, and filarioses) did occur at variable frequencies, but could be eliminated through the use of confirmatory testing.

Conclusions

Different metacestode-derived antigens of E. granulosus and E. multilocularis are valuable, widely accessible, and cost-efficient tools for the serological diagnosis of echinococcosis. However, confirmatory testing is necessary, due to the lack of species specificity and the occurrence of cross-reactions to other helminthic diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  1. Eckert J, Deplazes P, Kern P. Alveolar Echinococcosis (Echinococcus multilocularis) and other forms of echinococcosis (Echinococcus vogeli and Echinococcus oligarthrus). In: Brown D, Palmer SR, Torgerson PR, Soulsby EJL, editors. Zoonoses. Oxford: Oxford University Press; 2011. p. 671–701.

    Google Scholar 

  2. Brunetti E, Kern P, Vuitton DA, Writing Panel for the WHO Informal Working Group on Echinococcosis. Expert consensus for the diagnosis and treatment of cystic and alveolar echinococcosis in humans. Acta Trop. 2010;114:1–16.

    Article  PubMed  Google Scholar 

  3. Ammann RW, Eckert J. Cestodes. Echinococcus. Gastroenterol Clin North Am. 1996;25:655–89.

    Article  PubMed  CAS  Google Scholar 

  4. Torgerson PR, Schweiger A, Deplazes P, Pohar M, Reichen J, Ammann RW, et al. Alveolar echinococcosis: from a deadly disease to a well-controlled infection. Relative survival and economic analysis in Switzerland over the last 35 years. J Hepatol. 2008;49:72–7.

    Article  PubMed  Google Scholar 

  5. Eckert J, Deplazes P. Biological, epidemiological, and clinical aspects of echinococcosis, a zoonosis of increasing concern. Clin Microbiol Rev. 2004;17:107–35.

    Article  PubMed  Google Scholar 

  6. Schweiger A, Ammann RW, Candinas D, Clavien PA, Eckert J, Gottstein B, et al. Human alveolar echinococcosis after fox population increase, Switzerland. Emerg Infect Dis. 2007;13:878–82.

    PubMed  Google Scholar 

  7. Bruzinskaite R, Marcinkute A, Strupas K, Sokolovas V, Deplazes P, Mathis A, et al. Alveolar echinococcosis, Lithuania. Emerg Infect Dis. 2007;13:1618–9.

    PubMed  Google Scholar 

  8. Torgerson PR, Karaeva RR, Corkeri N, Abdyjaparov TA, Kuttubaev OT, Shaikenov BS. Human cystic echinococcosis in Kyrgystan: an epidemiological study. Acta Trop. 2003;85:51–61.

    Google Scholar 

  9. Vuitton DA, Zhou H, Bresson-Hadni S, Wang Q, Piarroux M, Raoul F, et al. Epidemiology of alveolar echinococcosis with particular reference to China and Europe. Parasitology. 2003;127:S87–107.

    Article  PubMed  Google Scholar 

  10. Ito A, Romig T, Takahashi K. Perspective on control options for Echinococcus multilocularis with particular reference to Japan. Parasitology. 2003;127:S159–72.

    Article  PubMed  Google Scholar 

  11. Sako Y, Nakao M, Nakaya K, Yamasaki H, Ito A. Recombinant antigens for serodiagnosis of cysticercosis and echinococcosis. Parasitol Int. 2006;55:S69–73.

    Article  PubMed  CAS  Google Scholar 

  12. Coudert J, Ambroise-Thomas P, Kien Truong T, Pothier MA. 1st results concerning the serologic diagnosis of hydatid cyst by a new technic of immunofluorescence on slides. Bull Soc Pathol Exot Filiales. 1967;60:555–63.

    PubMed  CAS  Google Scholar 

  13. Auer H, Hermentin K, Aspock H. Demonstration of a specific Echinococcus multilocularis antigen in the supernatant of in vitro maintained protoscolices. Zentralbl Bakteriol Mikrobiol Hyg A. 1988;268:416–23.

    PubMed  CAS  Google Scholar 

  14. Gottstein B, Eckert J, Fey H. Serological differentiation between Echinococcus granulosus and E. multilocularis infections in man. Z Parasitenkd. 1983;69:347–56.

    Article  PubMed  CAS  Google Scholar 

  15. Deplazes P, Gottstein B. A monoclonal antibody against Echinococcus multilocularis Em2 antigen. Parasitology. 1991;103:41–9.

    Article  PubMed  Google Scholar 

  16. Gottstein B, Deplazes P, Aubert M. Echinococcus multilocularis: immunological study on the “Em2-positive” laminated layer during in vitro and in vivo post-oncospheral and larval development. Parasitol Res. 1992;78:291–7.

    Article  PubMed  CAS  Google Scholar 

  17. Gottstein B, Eckert J, Michael SA, Thompson RCA. Echinococcus granulosus antigens: Immunoelectrophoretic and Western Blot analysis of hydatid cyst fluids. Parasitol Res. 1987;73:186–9.

    Article  PubMed  CAS  Google Scholar 

  18. Frayha GJ, Bahr GM, Haddad R. The lipids and phospholipids of hydatid protoscolices of Echinococcus granulosus (Cestoda). Int J Parasitol. 1980;10:213–6.

    Article  PubMed  CAS  Google Scholar 

  19. Frayha GJ, Haddad R. Comparative chemical composition of protoscolices and hydatid cyst fluid of Echinococcus granulosus (Cestoda). Int J Parasitol. 1980;10:359–64.

    Article  PubMed  CAS  Google Scholar 

  20. Lorenzo C, Ferreira HB, Monteiro KM, Rosenzvit M, Kamenetzky L, Garcia HH, et al. Comparative analysis of the diagnostic performance of six major Echinococcus granulosus antigens assessed in a double-blind, randomized multicenter study. J Clin Microbiol. 2005;43:2764–70.

    Article  PubMed  CAS  Google Scholar 

  21. MacPherson CNL, Vuitton DA, Gharbia HA, Caremani M, Frider B, Brunetti E, WHO informal working group on the development of a standardised classification for CE by US. International classification of ultrasound images in cystic echinococcosis for application in clinical and field epidemiological settings. Acta Trop. 2003;85:253–61.

    Google Scholar 

  22. Eckert J, Pohlenz J. On the effect of mebendazole on metacestodes of Mesocestoides corti and Echinococcus multilocularis (author’s translation). Tropenmed Parasitol. 1976;27:247–62.

    PubMed  CAS  Google Scholar 

  23. Muller N, Frei E, Nunez S, Gottstein B. Improved serodiagnosis of alveolar echinococcosis of humans using an in vitro-produced Echinococcus multilocularis antigen. Parasitology. 2007;134:879–88.

    Article  PubMed  CAS  Google Scholar 

  24. Oriol R, Williams JF, Perez Esandi MV, Oriol C. Purification of lipoprotein antigens of Echinococcus granulosus from sheep hydatid fluid. Am J Trop Med Hyg. 1971;20:569–74.

    PubMed  CAS  Google Scholar 

  25. Ortona E, Rigano R, Margutti P, Notargiacomo S, Ioppolo S, Vaccari S, et al. Native and recombinant antigens in the immunodiagnosis of human cystic echinococcosis. Parasite Immunol. 2000;22:553–9.

    Article  PubMed  CAS  Google Scholar 

  26. Ito A, Osawa Y, Nakao M, Horii T, Okamoto M, Itoh M, et al. Em18 and Em16, new serologic marker epitopes for alveolar echinococcosis in western blot analysis, are the only two epitopes recognized by commercially available weak positive (cut off) sera for Em2plus-ELISA. J Helminthol. 1995;69:369–71.

    Article  PubMed  CAS  Google Scholar 

  27. Ito A, Xiao N, Liance M, Sato MO, Sako Y, Mamuti W, et al. Evaluation of an enzyme-linked immunosorbent assay (ELISA) with affinity-purified Em18 and an ELISA with recombinant Em18 for differential diagnosis of alveolar echinococcosis: results of a blind test. J Clin Microbiol. 2002;40:4161–5.

    Article  PubMed  CAS  Google Scholar 

  28. Akobeng AK. Understanding diagnostic tests 3: receiver operating characteristic curves. Acta Paediatr. 2007;96:644–7.

    Article  PubMed  Google Scholar 

  29. Greiner M, Sohr D, Gobel P. A modified ROC analysis for the selection of cut-off values and the definition of intermediate results of serodiagnostic tests. J Immunol Methods. 1995;185:123–32.

    Article  PubMed  CAS  Google Scholar 

  30. Akobeng AK. Understanding diagnostic tests 2: likelihood ratios, pre- and post-test probabilities and their use in clinical practice. Acta Paediatr. 2007;96:487–91.

    Article  PubMed  Google Scholar 

  31. Leporrier J, Maurel J, Chiche L, Bara S, Segol P, Launoy G. A population-based study of the incidence, management and prognosis of hepatic metastases from colorectal cancer. Br J Surg. 2006;93:465–74.

    Article  PubMed  CAS  Google Scholar 

  32. Deplazes P, Grimm F, Sydler T, Tanner I, Kapel CM. Experimental alveolar echinococcosis in pigs, lesion development and serological follow up. Vet Parasitol. 2005;130:213–22.

    Article  PubMed  CAS  Google Scholar 

  33. Siracusano A, Bruschi F. Cystic echinococcosis: progress and limits in epidemiology and immunodiagnosis. Parassitologia. 2006;48:65–6.

    PubMed  CAS  Google Scholar 

  34. Ito A, Craig PS. Immunodiagnostic and molecular approaches for the detection of taeniid cestode infections. Trends Parasitol. 2003;19:377–81.

    Article  PubMed  CAS  Google Scholar 

  35. Siles-Lucas M, Gottstein B. Molecular tools for the diagnosis of cystic and alveolar echinococcosis. Trop Med Int Health. 2001;6:463–75.

    Article  PubMed  CAS  Google Scholar 

  36. Sanchez F, March F, Mercader M, Coll P, Munoz C, Prats G. Immunochemical localization of major hydatid fluid antigens in protoscoleces and cysts of Echinococcus granulosus from human origin. Parasite Immunol. 1991;13:583–92.

    Article  PubMed  CAS  Google Scholar 

  37. Gottstein B, Lengeler C, Bachmann P, Hagemann P, Kocher P, Brossard M, et al. Sero-epidemiological survey for alveolar echinococcosis (by Em2-ELISA) of blood donors in an endemic area of Switzerland. Trans R Soc Trop Med Hyg. 1987;81:960–4.

    Article  PubMed  CAS  Google Scholar 

  38. Craig PS, Giraudoux P, Shi D, Bartholomot B, Barnish G, Delattre P, et al. An epidemiological and ecological study of human alveolar echinococcosis transmission in south Gansu, China. Acta Trop. 2000;77:167–77.

    Article  PubMed  CAS  Google Scholar 

  39. Zhang W, Li J, McManus DP. Concepts in immunology and diagnosis of hydatid disease. Clin Microbiol Rev. 2003;16:18–36.

    Article  PubMed  CAS  Google Scholar 

  40. Rafiei A, Craig PS. The immunodiagnostic potential of protoscolex antigens in human cystic echinococcosis and the possible influence of parasite strain. Ann Trop Med Parasitol. 2002;96:383–9.

    Article  PubMed  CAS  Google Scholar 

  41. Rott MB, Fernandez V, Farias S, Ceni J, Ferreira HB, Haag KL, et al. Comparative analysis of two different subunits of antigen B from Echinococcus granulosus: gene sequences, expression in Escherichia coli and serological evaluation. Acta Trop. 2000;75:331–40.

    Article  PubMed  CAS  Google Scholar 

  42. Virginio VG, Hernandez A, Rott MB, Monteiro KM, Zandonai AF, Nieto A, et al. A set of recombinant antigens from Echinococcus granulosus with potential for use in the immunodiagnosis of human cystic hydatid disease. Clin Exp Immunol. 2003;132:309–15.

    Article  PubMed  CAS  Google Scholar 

  43. Jiang L, Wen H, Ito A. Immunodiagnostic differentiation of alveolar and cystic echinococcosis using ELISA test with 18-kDa antigen extracted from Echinococcus protoscoleces. Trans R Soc Trop Med Hyg. 2001;95:285–8.

    Article  PubMed  CAS  Google Scholar 

  44. Gottstein B, Jacquier P, Bresson-Hadni S, Eckert J. Improved primary immunodiagnosis of alveolar echinococcosis in humans by an enzyme-linked immunosorbent assay using the Em2plus antigen. J Clin Microbiol. 1993;31:373–6.

    PubMed  CAS  Google Scholar 

  45. Frosch PM, Frosch M, Pfister T, Schaad V, Bitter-Suermann D. Cloning and characterisation of an immunodominant major surface antigen of Echinococcus multilocularis. Mol Biochem Parasitol. 1991;48:121–30.

    Article  PubMed  CAS  Google Scholar 

  46. Xiao N, Mamuti W, Yamasaki H, Sako Y, Nakao M, Nakaya K, et al. Evaluation of use of recombinant Em18 and affinity-purified Em18 for serological differentiation of alveolar echinococcosis from cystic echinococcosis and other parasitic infections. J Clin Microbiol. 2003;41:3351–3.

    Article  PubMed  CAS  Google Scholar 

  47. Mamuti W, Yamasaki H, Sako Y, Nakao M, Xiao N, Nakaya K, et al. Molecular cloning, expression, and serological evaluation of an 8-kilodalton subunit of antigen B from Echinococcus multilocularis. J Clin Microbiol. 2004;42:1082–8.

    Article  PubMed  CAS  Google Scholar 

  48. Li T, Ito A, Chen X, Sako Y, Qiu J, Xiao N, et al. Specific IgG responses to recombinant antigen B and Em18 in cystic and alveolar echinococcosis in China. Clin Vaccine Immunol. 2010;17:470–5.

    Article  PubMed  CAS  Google Scholar 

  49. Tawfeek GM, Elwakil HS, El-Hoseiny L, Thabet HS, Sarhan RM, Awad NS, et al. Comparative analysis of the diagnostic performance of crude sheep hydatid cyst fluid, purified antigen B and its subunit (12 Kda), assessed by ELISA, in the diagnosis of human cystic echinococcosis. Parasitol Res. 2011;108:371–6.

    Article  PubMed  Google Scholar 

  50. Feng X, Wen H, Zhang Z, Chen X, Ma X, Zhang J, et al. Dot immunogold filtration assay (DIGFA) with multiple native antigens for rapid serodiagnosis of human cystic and alveolar echinococcosis. Acta Trop. 2010;113:114–20.

    Article  PubMed  CAS  Google Scholar 

  51. Czermak BV, Akhan O, Hiemetzberger R, Zelger B, Vogel W, Jaschke W, et al. Echinococcosis of the liver. Abdom Imaging. 2008;33:133–43.

    Article  PubMed  Google Scholar 

  52. Lim JH. Parasitic diseases in the abdomen: imaging findings. Abdom Imaging. 2008;33:130–2.

    Article  PubMed  Google Scholar 

  53. Espinoza JR, Maco V, Marcos L, Saez S, Neyra V, Terashima A, et al. Evaluation of Fas2-ELISA for the serological detection of Fasciola hepatica infection in humans. Am J Trop Med Hyg. 2007;76:977–82.

    PubMed  CAS  Google Scholar 

  54. Marcos LA, Terashima A, Gotuzzo E. Update on hepatobiliary flukes: fascioliasis, opisthorchiasis and clonorchiasis. Curr Opin Infect Dis. 2008;21:523–30.

    Article  PubMed  Google Scholar 

  55. Tsang VC, Brand JA, Boyer AE. An enzyme-linked immunoelectrotransfer blot assay and glycoprotein antigens for diagnosing human cysticercosis (Taenia solium). J Infect Dis. 1989;159:50–9.

    Article  PubMed  CAS  Google Scholar 

  56. Hubert K, Andriantsimahavandy A, Michault A, Frosch M, Muhlschlegel FA. Serological diagnosis of human cysticercosis by use of recombinant antigens from Taenia solium cysticerci. Clin Diagn Lab Immunol. 1999;6:479–82.

    PubMed  CAS  Google Scholar 

  57. Ito A, Plancarte A, Ma L, Kong Y, Flisser A, Cho SY, et al. Novel antigens for neurocysticercosis: simple method for preparation and evaluation for serodiagnosis. Am J Trop Med Hyg. 1998;59:291–4.

    PubMed  CAS  Google Scholar 

  58. Manzella A, Ohtomo K, Monzawa S, Lim JH. Schistosomiasis of the liver. Abdom Imaging. 2008;33:7.

    Article  Google Scholar 

  59. Ross AG, Bartley PB, Sleigh AC, Olds GR, Li Y, Williams GM, et al. Schistosomiasis. N Engl J Med. 2002;346:1212–20.

    Article  PubMed  Google Scholar 

  60. Eckert J, Gemmel MA, Meslin F-X, Pawlowski ZS. WHO/OIE manual on echinococcosis in humans and animals: a public health problem of global concern. Paris: World Organisation for Animal Health and World Health Organization; 2001.

    Google Scholar 

  61. Torgerson PR, Deplazes P. Echinococcosis: diagnosis and diagnostic interpretation in population studies. Trends Parasitol. 2009;25:164–70.

    Article  PubMed  Google Scholar 

  62. Reuter S, Nussle K, Kolokythas O, Haug U, Rieber A, Kern P, et al. Alveolar liver echinococcosis: a comparative study of three imaging techniques. Infection. 2001;29:119–25.

    Article  PubMed  CAS  Google Scholar 

Download references

Conflict of interest

None.

Author information

Authors and Affiliations

  1. Institute of Parasitology, University of Zurich, Winterthurerstrasse 266a, 8057, Zurich, Switzerland

    A. Schweiger, F. Grimm, I. Tanner & P. Deplazes

  2. Division of Gastroenterology and Hepatology, University Hospital of Zurich, Zurich, Switzerland

    B. Müllhaupt & K. Bertogg

  3. Institute of Parasitology, University of Bern, Bern, Switzerland

    N. Müller

Authors
  1. A. Schweiger
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F. Grimm
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. I. Tanner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B. Müllhaupt
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K. Bertogg
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. N. Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. P. Deplazes
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to F. Grimm.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schweiger, A., Grimm, F., Tanner, I. et al. Serological diagnosis of echinococcosis: the diagnostic potential of native antigens. Infection 40, 139–152 (2012). https://doi.org/10.1007/s15010-011-0205-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s15010-011-0205-6

Keywords

Search

Navigation