Skip to main content

Detection of toxoplasma-specific immunoglobulin G in human sera: performance comparison of in house Dot-ELISA with ECLIA and ELISA

European Journal of Clinical Microbiology & Infectious Diseases volume 37, pages 1421–1429 (2018)Cite this article

Abstract

In the current study, performance of electrochemiluminescence immunoassay (ECLIA) in detection of anti-toxoplasma IgG in human sera was compared with that of enzyme-linked immunosorbent assay (ELISA). Furthermore, performance of an in house Dot-ELISA in detection of anti-toxoplasma IgG was compared with that of ECLIA and ELISA. In total, 219 human sera were tested to detect anti-toxoplasma IgG using Dynex DS2® and Roche Cobas® e411 Automated Analyzers. Discordant results rechecked using immunofluorescence assay (IFA). Then, sera were used in an in house Dot-ELISA to assess toxoplasma-specific IgG. Of the 219 samples, two samples were found undetermined using ECLIA but reactive using ELISA. Using IFA, the two sera were reported unreactive. Furthermore, two samples were found reactive using ECLIA and unreactive using ELISA. These samples were reported reactive using IFA. The overall agreement for the two former methods was 98% (rZ0.98.1; P < 0.001). The intrinsic parameters calculated for in house Dot-ELISA included sensitivity of 79.5, specificity of 78.2, and accuracy of 78.9%, compared to ECLIA and ELISA. Positive and negative predictive values included 82.9 and 74.2%, respectively. A 100% sensitivity was found in in house Dot-ELISA for highly reactive sera in ECLIA and ELISA. ECLIA is appropriate for the first-line serological screening tests and can replace ELISA due to high speed, sensitivity, and specificity, particularly in large laboratories. Dot-ELISA is a rapid, sensitive, specific, cost-effective, user-friendly, and field-portable technique and hence can be used for screening toxoplasmosis, especially in rural fields or less equipped laboratories.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Abbreviations

ECLIA:

electrochemiluminescence immunoassay

ELISA:

enzyme-linked immunosorbent assay

IgG:

immunoglobulin G

DT:

dye test

IFA:

immunofluorescence assay

CFT:

complement fixation test

MAT:

modified agglutination test

LAT:

latex agglutination test

DAT:

direct agglutination test

IFAT:

indirect fluorescent antibody test

IHA:

indirect hemagglutination assay

PBS-M 3%:

phosphate-buffered saline containing 3% of non-fat dry milk

PBS-T:

phosphate-buffered saline containing tween 20

HRP:

horseradish peroxidase

OPD:

ortho phenylenediamine

OD:

optical density

NC:

nitrocellulose membrane

IBM:

International Business Machines Corporation

SPSS:

Statistical Package for the Social Sciences

References

  1. Dubey JP (2010). Toxoplasmosis of animals and humans. 2rd edn. CRC Press Inc. Boca Raton, New York, pp 1–313

  2. Alvarado-Esquirel C, Estrada-Martinez S, Liesenfeld O (2011) Toxoplasma gondii infection in workers occupationally exposed to unwashed raw fruits and vegetables: a case control seroprevalence study. Parasit Vectors 16:4–235

    Google Scholar 

  3. Tenter AM, Heckeroth AR, Weiss LM (2000) Toxoplasma gondii: from animals to humans. Int J Parasitol 30:1217–1258

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Walle F, Kebede N, Tsegaye A, Kassa T (2013) Seroprevalence and risk factors for toxoplasmosis in HIV infected and non-infected individuals in Bahir Dar, Northwest Ethiopia. Parasit Vectors 6:15–10

    Article  PubMed  PubMed Central  Google Scholar 

  5. Tuazon CU (1989) Toxoplasmosis in AIDS patients. J Antimicrob Chemother 23(suppl A):77–82

    Article  PubMed  Google Scholar 

  6. Robert-Gangneux F, Darde ML (2012) Epidemiology of and diagnostic strategies for toxoplasmosis. Clin Microbiol Rev 25:264–296

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  7. Liu Q, Wang Z-D, Huang S-Y, Zhu X-Q (2015) Diagnosis of toxoplasmosis and typing of Toxoplasma gondii. Parasit Vectors 8(1):292

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  8. Acharya D, Bastola P, Le L, Paul AM, Fernandez E, Diamond MS et al (2016) An ultrasensitive electrogenerated chemiluminescence-based immunoassay for specific detection of Zika virus. Sci Rep 2016(6):32227

    Article  CAS  Google Scholar 

  9. Dard C, Fricker-Hidalgo H, Brenier-Pinchart MP, Pelloux H (2016) Relevance of and new developments in serology for toxoplasmosis. Trends Parasitol 32:492–506

    Article  PubMed  Google Scholar 

  10. Pappas MG, Hajkowski R, Hockmeyer WT (1983) Dot enzyme-linked immunosorbent assay (Dot-ELISA): a micro technique for the rapid diagnosis of visceral leishmaniasis. J lmmlmologcal Methods 64:205–214

    Article  CAS  Google Scholar 

  11. Gupta A, Dixit AK, Dixit P (2008) Dot-ELISA in the diagnosis of parasitic diseases of animals. J Parasit Dis 32(1):10–14

    Google Scholar 

  12. Zhang K, Lin G, Han Y, Li J (2016) Serological diagnosis of toxoplasmosis and standardization. Clin Chim Acta 46:83–89

    Article  CAS  Google Scholar 

  13. Marquette CA, Blum LJ (2008) Electro-chemiluminescent biosensing. Anal Bioanal Chem 390:155–168

    Article  PubMed  CAS  Google Scholar 

  14. Ali-Heydari S, Keshavarz H, Shojaee S, Mohebali M (2013) Diagnosis of antigenic markers of acute toxoplasmosis by IgG avidity immunoblotting. Parasite 20(18):20–18

    Google Scholar 

  15. Hawkes R, Niday E, Gordon J (1982) A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem 119(1):142–147

    Article  PubMed  CAS  Google Scholar 

  16. Pappas MG (1988) Recent applications of the Dot-ELISA in immunoparasitology. Vet Parasitol 29(2–3):105–129

    Article  PubMed  CAS  Google Scholar 

  17. IBM Corp Released (2012) IBM SPSS statistics for windows, version 21.0. Armonk, NY: IBM Corp

  18. Landis JR, Koch GG (1977) The measurement of observer agreement for categorical data. Biometrics 33(1):159–174

    Article  PubMed  CAS  Google Scholar 

  19. Jacobson RH (1998) Validation of serological assays for diagnosis of infectious diseases. Rev Sci Tech 17(2):469–526

    Article  PubMed  CAS  Google Scholar 

  20. Hayde M, Salzer HR, Gittler G, Aspock H, Pollak A (1995) Microparticle enzyme immunoassay (MEIA) for toxoplasma specific immunoglobulin G in comparison to the Sabin-Feldman dye test. A pilot study. Wien Klin Wochenschr 107:133–136

    PubMed  CAS  Google Scholar 

  21. Kasper DC, Prusa AR, Hayde M, Gerstl N, Pollak A, Herkner KR, Reiter-Reisacher R (2009) Evaluation of the Vitros ECiQ immunodiagnostic system for detection of anti-toxoplasma immunoglobulin G and immunoglobulin M antibodies for confirmatory testing for acute Toxoplasma gondii infection in pregnant women. J Clin Microbiol 47:164–167

    Article  PubMed  CAS  Google Scholar 

  22. Petersen E, Borobio MV, El G et al (2005) European multicenter study of the LIAISON automated diagnostic system for determination of Toxoplasma gondii-specific immunoglobulin G (IgG) and IgM and the IgG avidity index. J Clin Microbiol 43:1570–1574

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Tekkesin N, Keskin K, Kılınc C, Orgen N, Molo K (2011) Detection of immunoglobulin G antibodies to Toxoplasma gondii: evaluation of two commercial immunoassay systems. J Microbiol Immunol Infect 44:21–26

    Article  PubMed  CAS  Google Scholar 

  24. Prusa AR, Hayde M, Unterasinger L, Pollak A, Herkner KR et al (2010) Evaluation of the Roche Elecsys Toxo IgG and IgM electrochemiluminescence immunoassay for the detection of gestational Toxoplasma infection. Diagn Microbiol Infect Dis 68(4):352–357

    Article  PubMed  CAS  Google Scholar 

  25. Murat JB, Hidalgo HF, Brenier-Pinchart MP, Pelloux H (2013) Human toxoplasmosis: which biological diagnostic tests are best suited to which clinical situations? Expert Rev Anti-Infect Ther 11(9):943–956

    Article  PubMed  CAS  Google Scholar 

  26. Wilson M, Remington JS, Clavet C, Varney G, Press C, Ware D (1997) Evaluation of six commercial kits for detection of human immunoglobulin M antibodies to Toxoplasma gondii. J Clin Microbiol 35:3112–3115

    PubMed  PubMed Central  CAS  Google Scholar 

  27. Syme NR, Toft AD, Stoddart M, Beckett GJ (2011) Clinical performance of the Roche cobas e411 automated assay system for thyrotropin-receptor antibodies for the diagnosis of Graves’ disease. Ann Clin Biochem 48(5):471–473

    Article  PubMed  CAS  Google Scholar 

  28. Selseleh M, Keshavarz H, Mohebali M, Shojaee S, et al (2012) Production and Evaluation of Toxoplasma gondii Recombinant GRA7 for Serodiagnosis of Human Infections. Korean J Parasitol 50(3):233–238

  29. Leser PG, de Assis Rocha LS, Moura MEG, Ferreira AW (2003) Comparison of semi-automatized assays for anti-T. gondii IgG detection in low-reactivity serum samples: importance of the results in patient counseling. J Bras Patol Med Lab 39:107–110

    Article  CAS  Google Scholar 

  30. Meylan P, Paris L, Liesenfeld O (2015) Multicenter evaluation of the Elecsys Toxo IgG and IgM tests for the diagnosis of infection with Toxoplasma gondii. Eur J Microbiol Immunol 5(2):150–158

    Article  CAS  Google Scholar 

  31. Prusa AR, Hayde M, Unterasinger L, Pollak A, Herkner KR et al (2010) Evaluation of the Roche Elecsys Toxo IgG and IgM electrochemiluminescence immunoassay for the detection of gestational Toxoplasma infection. Diagn Microbiol Infect Dis 68:352–357

    Article  PubMed  CAS  Google Scholar 

  32. Van Helden J (2009) Performance of Elecsys toxo IgG and IgM immunoassays. Clin Lab 55:267–273

    PubMed  Google Scholar 

  33. Murat JB, L’Ollivier C, Fricker Hidalgo H, Franck J, Pelloux H et al (2012) Evaluation of the new Elecsys Toxo IgG avidity assay for toxoplasmosis and new insights into the interpretation of avidity results. Clin Vaccine Immunol 19:1838–1843

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  34. Schaefer LE, Dyke JW, Meglio FD, Murray PR, Crafts W, Niles AC (1989) Evaluation of microparticle enzyme immunoassays for immunoglobulins G and M to rubella virus and Toxoplasma gondii on the Abbott IMx automated analyzer. J Clin Microbiol 27:2410–2413

    PubMed  PubMed Central  CAS  Google Scholar 

  35. Selseleh M, Keshavarz H, Mohebali M, Shojaee S, Modarressi MH, Eshragian M (2012) Production and evaluation of Toxoplasma gondii recombinant surface antigen 1 (SAG1) for serodiagnosis of acute and chronic Toxoplasma infection in human sera. Iran J Parasitol 7:1–9

  36. Tenter AM (1988) Comparison of Dot-ELISA, ELISA and IFAT for the detection of IgG antibodies to Sarcocystis muris in experimentally infected and immunized mice. Vet Parasit 29:89–104

    Article  CAS  Google Scholar 

  37. Vercammen F, Berkvens D, Brandt J, Vansteenkiste W (1998) A sensitive and specific 30-min Dot-ELISA for the detection of anti-leishmania antibodies in the dog. Vet Parasitol 79:221–228

    Article  PubMed  CAS  Google Scholar 

  38. Wanduragala L, Kakoma I, Clabaugh GW, Abeygunawardena I, Levy MG, Ristic M (1987) Development of dot-enzyme immunoassay for diagnosis of canine babesiosis. Am J Trop Med Hyg 36:20–21

    Article  PubMed  CAS  Google Scholar 

  39. Montenegro James S, Guillen AT, Toro M (1992) Dot-ELISA para el diagnostico serologico de la anaplasmosis y babesiosis bovina. Rev Ciesstifica FCV de Luz II:23–29

  40. Camus E, Montenegro James S (1994) Bovine anaplasmosis and babesiosis in the lesser antilles: risk assessment of an unstable epidemiologic situation. Vet Res 25:313–317

    PubMed  CAS  Google Scholar 

  41. Mattioli RC, Janneh L, Corr N, Faye JA, Pandey US, Verhulet A (1997) Seasonal prevalence of ticks and tick transmitted haemoparasites in traditionally managed N’Dama cattle with reference to strategic tick control in the Gambia. Med Vet Entomol 11:342–348

    Article  PubMed  CAS  Google Scholar 

  42. Jafarpour Azami S, Keshavarz H, Rezaian M, Mohebali M, Shojaee S (2011) Rapid detection of Toxoplasma gondii antigen in experimentally infected mice by dot-ELISA. Iranian J Parasitol 6:28–33

    CAS  Google Scholar 

  43. Siavashi MR, Taherkhani H, Rezaei KR, Razavi Deligani MR, Assmar M (2005) Comparison of dot-ELISA and sandwich ELISA diagnostic tests in detection of human hydatidosis. Iran Biomed J 9:91–94

    Google Scholar 

Download references

Acknowledgements

We would like to acknowledge all staff from the toxoplasmosis laboratory (Department of Medical Parasitology and Mycology, Tehran University of Medical Sciences, Tehran, Iran) for their useful collaboration.

Funding

This study was supported by the Tehran University of Medical Sciences and Health Services (Project Number: 29999-160-03-94), Tehran, Iran.

Author information

Affiliations

  1. Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Pour Sina Street, Ghods Avenue, Enghelab Avenue, P.O. Box: 1417613191, Tehran, Iran

    Aref Teimouri, Saeedeh Shojaee, Mehdi Mohebali, Nima Zouei, Mostafa Rezaian & Hossein Keshavarz

  2. Students Scientific Research Center, Tehran University of Medical Sciences, Tehran, Iran

    Aref Teimouri

  3. Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Mohammad Hossein Modarressi

Authors
  1. Aref Teimouri
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mohammad Hossein Modarressi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Saeedeh Shojaee
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mehdi Mohebali
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nima Zouei
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mostafa Rezaian
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hossein Keshavarz
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

HK and MHM conceived the study. NZ collected and prepared the samples. AT performed the experiments with substantial contributions by SS. AT drafted the manuscript and revised by SS, MM, and MR. All authors read and approved the final version of the manuscript.

Corresponding author

Correspondence to Hossein Keshavarz.

Ethics declarations

Ethics approval and consent to participate

The study was approved by the Ethical Committee of Tehran University of Medical Sciences.

Consent for publication

Not applicable (no individual person’s data).

Availability of data and material

The data that support the findings of this study are available on reasonable request to the corresponding author.

Competing interests

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Teimouri, A., Modarressi, M.H., Shojaee, S. et al. Detection of toxoplasma-specific immunoglobulin G in human sera: performance comparison of in house Dot-ELISA with ECLIA and ELISA. Eur J Clin Microbiol Infect Dis 37, 1421–1429 (2018). https://doi.org/10.1007/s10096-018-3266-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10096-018-3266-y

Keywords

  • Anti-toxoplasma IgG
  • In house Dot-ELISA
  • ECLIA
  • ELISA