P35 and P22 Toxoplasma gondii antigens abbreviate regions to diagnose acquired toxoplasmosis during pregnancy: toward single-sample assays
-
Juan G. Costa
and
Iván S. Marcipar
Abstract
Background:
P35 and P22 Toxoplasma gondii proteins are recognized by specific IgG at the early infection stage, making them ideal for acute toxoplasmosis pregnancy control. Both proteins have been studied to discriminate between acute and chronic toxoplasmosis. However, results were hardly comparable because different protein obtainment procedures led to different antigens, the reference panels used were not optimally typified, and avidity tests were either not performed or narrowly examined.
Methods:
We bioinformatically predicted P35 and P22 regions with the highest density of epitopes, and expressed them in pET32/BL21DE3 alternative expression system, obtaining the soluble proteins rP35a and rP22a. We assessed their diagnostic performance using pregnant woman serum samples typified as: not infected, NI (IgG−, IgM−), typical-chronic, TC (IgM−, IgG+), presumably acute, A (IgG+, IgM+, low-avidity IgG), and recently chronic, RC (IgG+, IgM+, high-avidity IgG).
Results:
rP35a performed better than rP22a to differentiate A from RC, the areas under the curve (AUC) being 0.911 and 0.818, respectively. They, however, performed similarly to differentiate A from TC+RC (AUC: 0.915 and 0.907, respectively). rP35a and rP22a evaluation by avidity ELISA to discriminate A from RC rendered AUC values of 0.974 and 0.921, respectively. The indirect ELISA and avidity ELISA results analyzed in tandem were consistent with those obtained using commercial kits.
Conclusions:
rP35a and rP22a features suggest that, with complementary use, they could replace parasite lysate for toxoplasmosis infection screening and for acute toxoplasmosis diagnosis. Our proposal should be validated by a longitudinal study and may lead to a reliable toxoplasmosis pregnancy control, performing tests in only one serum sample.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentinean Research Council, supported this work (PIP N° 112 2009 0100545).
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
References
1. Sensini A. Toxoplasma gondii infection in pregnancy: opportunities and pitfalls of serological diagnosis. Clin Microbiol Infect 2006;12:504–12.10.1111/j.1469-0691.2006.01444.xSearch in Google Scholar
2. Montoya JG, Remington JS. Management of Toxoplasma gondii infection during pregnancy. Clin Infect Dis 2008;47:554–66.10.1086/590149Search in Google Scholar
3. De Paschale M, Agrappi C, Belvisi L, Cagnin D, Cerulli T, Clerici P, et al. Revision of the positive predictive value of IgM anti-Toxoplasma antibodies as an index of recent infection. New Microbiol 2008;31:105–11.Search in Google Scholar
4. Candolfi E, Pastor R, Huber R, Filisetti D, Villard O. IgG avidity assay firms up the diagnosis of acute toxoplasmosis on the first serum sample in immunocompetent pregnant women. Diagn Microbiol Infect Dis 2007;58:83–8.10.1016/j.diagmicrobio.2006.12.010Search in Google Scholar
5. Pelloux H, Brun E, Vernet G, Marcillat S, Jolivet M, Guergour D, et al. Determination of anti-Toxoplasma gondii immunoglobulin G avidity: adaptation to the Vidas system (bioMerieux). Diagn Microbiol Infect Dis 1998;32:69–73.10.1016/S0732-8893(98)00077-7Search in Google Scholar
6. Villard O, Breit L, Cimon B, Franck J, Fricker-Hidalgo H, Godineau N, et al. Comparison of four commercially available avidity tests for Toxoplasma gondii-specific IgG antibodies. Clin Vaccine Immunol 2013;20:197–204.10.1128/CVI.00356-12Search in Google Scholar PubMed PubMed Central
7. Parmley SF, Sgarlato GD, Mark J, Prince JB, Remington JS. Expression, characterization, and serologic reactivity of recombinant surface antigen P22 of Toxoplasma gondii. J Clin Microbiol 1992;30:1127–33.10.1128/jcm.30.5.1127-1133.1992Search in Google Scholar PubMed PubMed Central
8. Golkar M, Azadmanesh K, Khalili G, Khoshkholgh-Sima B, Babaie J, Mercier C, et al. Serodiagnosis of recently acquired Toxoplasma gondii infection in pregnant women using enzyme-linked immunosorbent assays with a recombinant dense granule GRA6 protein. Diagn Microbiol Infect Dis 2008;61:31–9.10.1016/j.diagmicrobio.2007.09.003Search in Google Scholar PubMed
9. Martin V, Supanitsky A, Echeverria PC, Litwin S, Tanos T, De Roodt AR, et al. Recombinant GRA4 or ROP2 protein combined with alum or the gra4 gene provides partial protection in chronic murine models of toxoplasmosis. Clin Diagn Lab Immunol 2004;11:704–10.10.1128/CDLI.11.4.704-710.2004Search in Google Scholar PubMed PubMed Central
10. Redlich A,Muller WA. Serodiagnosis of acute toxoplasmosis using a recombinant form of the dense granule antigen GRA6 in an enzyme-linked immunosorbent assay. Parasitol Res 1998;84:700–6.10.1007/s004360050473Search in Google Scholar PubMed
11. Li S, Galvan G, Araujo FG, Suzuki Y, Remington JS, Parmley S. Serodiagnosis of recently acquired Toxoplasma gondii infection using an enzyme-linked immunosorbent assay with a combination of recombinant antigens. Clin Diagn Lab Immunol 2000;7:781–7.10.1128/CDLI.7.5.781-787.2000Search in Google Scholar PubMed PubMed Central
12. Lu B, Wu S, Shi Y, Zhang R, Zou L, Gao S, et al. Toxoplasma gondii: expression pattern and detection of infection using full-length recombinant P35 antigen. Exp Parasitol 2006;113:83–90.10.1016/j.exppara.2005.12.014Search in Google Scholar PubMed
13. Aubert D, Maine GT, Villena I, Hunt JC, Howard L, Sheu M, et al. Recombinant antigens to detect Toxoplasma gondii-specific immunoglobulin G and immunoglobulin M in human sera by enzyme immunoassay. J Clin Microbiol 2000;38:1144–50.10.1128/JCM.38.3.1144-1150.2000Search in Google Scholar PubMed PubMed Central
14. Hiszczyñska-Sawicka E, Kur J, Pietkiewicz H, Holec L, Gasior A, Myjak P. Efficient production of the Toxoplasma gondii GRA6, p35 and SAG2 recombinant antigens and their applications in the serodiagnosis of toxoplasmosis. Acta Parasitol 2005;50:249–54.Search in Google Scholar
15. Bela SR, Oliveira Silva DA, Cunha-Junior JP, Pirovani CP, Chaves-Borges FA, Reis de CF, et al. Use of SAG2A recombinant Toxoplasma gondii surface antigen as a diagnostic marker for human acute toxoplasmosis: analysis of titers and avidity of IgG and IgG1 antibodies. Diagn Microbiol Infect Dis 2008;62:245–54.10.1016/j.diagmicrobio.2008.05.017Search in Google Scholar PubMed
16. Pfrepper KI, Enders G, Gohl M, Krczal D, Hlobil H, Wassenberg D, et al. Seroreactivity to and avidity for recombinant antigens in toxoplasmosis. Clin Diagn Lab Immunol 2005;12:977–82.10.1128/CDLI.12.8.977-982.2005Search in Google Scholar PubMed PubMed Central
17. Marcipar IS, Olivares ML, Robles L, Dekanty A, Marcipar A, Silber AM. The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2beta protein is influenced by its expression system. Protein Expr Purif 2004;34:1–7.10.1016/j.pep.2003.11.022Search in Google Scholar PubMed
18. Beghetto E, Buffolano W, Spadoni A, Del PM, Di CM, Minenkova O, et al. Use of an immunoglobulin G avidity assay based on recombinant antigens for diagnosis of primary Toxoplasma gondii infection during pregnancy. J Clin Microbiol 2003;41:5414–8.10.1128/JCM.41.12.5414-5418.2003Search in Google Scholar PubMed PubMed Central
19. Pietkiewicz H, Hiszczynska-Sawicka E, Kur J, Petersen E, Nielsen HV, Paul M, et al. Usefulness of Toxoplasma gondii recombinant antigens (GRA1, GRA7 and SAG1) in an immunoglobulin G avidity test for the serodiagnosis of toxoplasmosis. Parasitol Res 2007;100:333–7.10.1007/s00436-006-0265-1Search in Google Scholar PubMed
20. Elyasi H, Babaie J, Fricker-Hidalgo H, Brenier-Pinchart MP, Zare M, Sadeghiani G, et al. Use of dense granule antigen GRA6 in an immunoglobulin G avidity test to exclude acute Toxoplasma gondii infection during pregnancy. Clin Vaccine Immunol 2010;17:1349–55.10.1128/CVI.00199-10Search in Google Scholar PubMed PubMed Central
21. Holec-Gasior L, Ferra B, Hiszczynska-Sawicka E, Kur J. The optimal mixture of Toxoplasma gondii recombinant antigens (GRA1, P22, ROP1) for diagnosis of ovine toxoplasmosis. Vet Parasitol 2014;206:146–52.10.1016/j.vetpar.2014.09.018Search in Google Scholar PubMed
22. Deshpande PS, Kotresha D, Noordin R, Yunus MH, Saadatnia G, Golkar M, et al. IgG avidity Western blot using Toxoplasma gondii rGRA-7 cloned from nucleotides 39-711 for serodiagnosis of acute toxoplasmosis. Rev Inst Med Trop Sao Paulo 2013;55:79–83.10.1590/S0036-46652013000200003Search in Google Scholar
23. Makino M, Uemura N, Moroda M, Kikumura A, Piao LX, Mohamed RM, et al. Innate immunity in DNA vaccine with Toxoplasma gondii-heat shock protein 70 gene that induces DC activation and Th1 polarization. Vaccine 2011;29:1899–905.10.1016/j.vaccine.2010.12.118Search in Google Scholar PubMed
24. Costa JG, Faccendini PL, Sferco SJ, Lagier CM, Marcipar IS. Evaluation and comparison of the ability of online available prediction programs to predict true linear B-cell epitopes. Protein Pept Lett 2013;20:724–30.10.2174/0929866511320060011Search in Google Scholar PubMed
25. Kringelum JV, Lundegaard C, Lund O, Nielsen M. Reliable B cell epitope predictions: impacts of method development and improved benchmarking. PLoS Comput Biol 2012;8:e1002829.10.1371/journal.pcbi.1002829Search in Google Scholar PubMed PubMed Central
26. Babaie J, Miri M, Sadeghiani G, Zare M, Khalili G, Golkar M. Expression and single-step purification of GRA8 antigen of Toxoplasma gondii in Escherichia coli. Avicenna J Med Biotechnol 2011;3:67–77.Search in Google Scholar
27. Khanaliha K, Motazedian MH, Kazemi B, Shahriari B, Bandehpour M, Sharifniya Z. Evaluation of recombinant SAG1, SAG2, and SAG3 antigens for serodiagnosis of toxoplasmosis. Korean J Parasitol 2014;52:137–42.10.3347/kjp.2014.52.2.137Search in Google Scholar PubMed PubMed Central
28. Singh H, Tewari AK, Mishra AK, Maharana B, Sudan V, Raina OK, et al. Detection of antibodies to Toxoplasma gondii in domesticated ruminants by recombinant truncated SAG2 enzyme-linked immunosorbent assay. Trop Anim Health Prod 2015;47:171–8.10.1007/s11250-014-0703-5Search in Google Scholar PubMed
29. Roberts A, Hedman K, Luyasu V, Zufferey J, Bessieres MH, Blatz RM, et al. Multicenter evaluation of strategies for serodiagnosis of primary infection with Toxoplasma gondii. Eur J Clin Microbiol Infect Dis 2001;20:467–74.10.1007/PL00011289Search in Google Scholar
30. Drapala D, Holec-Gasior L, Kur J, Ferra B, Hiszczynska-Sawicka E, Lautenbach D. A new human IgG avidity test, using mixtures of recombinant antigens (rROP1, rSAG2, rGRA6), for the diagnosis of difficult-to-identify phases of toxoplasmosis. Diagn Microbiol Infect Dis 2014;79:342–6.10.1016/j.diagmicrobio.2014.03.005Search in Google Scholar PubMed
©2017 Walter de Gruyter GmbH, Berlin/Boston
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- Opinion Papers
- Improving quality in the preanalytical phase through innovation, on behalf of the European Federation for Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for Preanalytical Phase (WG-PRE)
- Metabolite profiling can change health-care delivery to obese patients with fatty liver disease: the search for biomarkers
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- Rapid screening for targeted genetic variants via high-resolution melting curve analysis
- Evaluation of the new cobas® HCV genotyping test based on real-time PCRs of three different HCV genome regions
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- Spanish Preanalytical Quality Monitoring Program (SEQC), an overview of 12 years’ experience
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- Prevalence of pseudonatremia in a clinical laboratory – role of the water content
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- Performance evaluation of ImmunoCAP® ISAC 112: a multi-site study
- Clinical autoantibody detection by microarray
- Cardiovascular Diseases
- Kinetics of troponin I in patients with myocardial injury after noncardiac surgery
- Infectious Diseases
- P35 and P22 Toxoplasma gondii antigens abbreviate regions to diagnose acquired toxoplasmosis during pregnancy: toward single-sample assays
- Letters to the Editor
- Deciphering a macro-troponin I complex; a case report
- Interference of laboratory disinfection with trichloro-isocyanuric acid on cardiac troponin I measurement using the Vitros immunoassay system
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- Congress Abstracts
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Articles in the same Issue
- Frontmatter
- Editorial
- The central role of external quality assurance in harmonisation and standardisation for laboratory medicine
- Review
- Fecal calprotectin in inflammatory bowel diseases: update and perspectives
- Mini Review
- Factor VIIa-antithrombin complex: a possible new biomarker for activated coagulation
- Opinion Papers
- Improving quality in the preanalytical phase through innovation, on behalf of the European Federation for Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for Preanalytical Phase (WG-PRE)
- Metabolite profiling can change health-care delivery to obese patients with fatty liver disease: the search for biomarkers
- Genetics and Molecular Diagnostics
- Rapid screening for targeted genetic variants via high-resolution melting curve analysis
- Evaluation of the new cobas® HCV genotyping test based on real-time PCRs of three different HCV genome regions
- General Clinical Chemistry and Laboratory Medicine
- Harmonisation of serum dihydrotestosterone analysis: establishment of an external quality assurance program
- Spanish Preanalytical Quality Monitoring Program (SEQC), an overview of 12 years’ experience
- Peer groups splitting in Croatian EQA scheme: a trade-off between homogeneity and sample size number
- Prevalence of pseudonatremia in a clinical laboratory – role of the water content
- Retrospective validation of a β-trace protein interpretation algorithm for the diagnosis of cerebrospinal fluid leakage
- Economic evaluation of procalcitonin-guided antibiotic therapy in acute respiratory infections: a Chinese hospital system perspective
- Performance evaluation of ImmunoCAP® ISAC 112: a multi-site study
- Clinical autoantibody detection by microarray
- Cardiovascular Diseases
- Kinetics of troponin I in patients with myocardial injury after noncardiac surgery
- Infectious Diseases
- P35 and P22 Toxoplasma gondii antigens abbreviate regions to diagnose acquired toxoplasmosis during pregnancy: toward single-sample assays
- Letters to the Editor
- Deciphering a macro-troponin I complex; a case report
- Interference of laboratory disinfection with trichloro-isocyanuric acid on cardiac troponin I measurement using the Vitros immunoassay system
- Massive interference in free T4 and free T3 assays misleading clinical judgment
- In defense of aldosterone measurement by immunoassay: a broader perspective
- The prevalence of hemolysis – a survey using hemolysis index
- Detection of BRAFV600K mutant tumor-derived DNA in the pleural effusion from a patient with metastatic melanoma
- Evaluation of the accuracy of the Greiner Bio-One FC Mix Glucose tube
- Congress Abstracts
- 4th EFLM-BD European Conference on Preanalytical Phase Amsterdam (NL), 24–25 March 2017
