Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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A New IgG Immunoblot Kit for Diagnosis of Toxoplasmosis in Pregnant Women

  • Khammari, Imen (Research Unit, UR12SP34, Farhat Hached University Hospital) ;
  • Saghrouni, Fatma (Research Unit, UR12SP34, Farhat Hached University Hospital) ;
  • Lakhal, Sami (Research Laboratory of Medical Parasitology, Biotechnology and Biomolecules, LR 11 IPT06 Pasteur Institute of Tunis) ;
  • Bouratbine, Aida (Research Laboratory of Medical Parasitology, Biotechnology and Biomolecules, LR 11 IPT06 Pasteur Institute of Tunis) ;
  • Said, Moncef Ben (Research Unit, UR12SP34, Farhat Hached University Hospital) ;
  • Boukadida, Jalel (Research Unit, UR12SP34, Farhat Hached University Hospital)
  • Received : 2014.04.23
  • Accepted : 2014.06.15
  • Published : 2014.10.31
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Abstract

The determination of the accurate immune status of pregnant women is crucial in order to prevent congenital toxoplasmosis. Equivocal results with conventional serological techniques are not uncommon when IgG titers are close to the cut-off value of the test, so that a confirmatory technique is needed. For this purpose, we developed a homemade immunoblot (IB) using soluble extract of Toxoplasma gondii tachyzoites and assessed it by testing 154 positive, 100 negative, and 123 equivocal sera obtained from pregnant women. In order to select the more valuable bands in terms of sensitivity and specificity, we used the Youden Index (YI). The highest YIs were those given by the 32, 36, 98, 21, and 33 bands. The simultaneous presence on the same blot of at least 3 bands showed a much higher YI (0.964) and was adapted as the positivity criterion. The analysis of results showed that our homemade IB correlated well with the commercial LDBIO Toxo II $IgG^{(R)}$ kit recently recommended as a confirmatory test (96.7% of concordance).

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References

  1. Dunn D, Wallon M, Peyron F, Petersen E, Peckham C, Gilbert R. Mother-to-child transmission of toxoplasmosis: risk estimates for clinical counselling. Lancet 1999; 353: 1829-1833. https://doi.org/10.1016/S0140-6736(98)08220-8
  2. Flori P, Chene G, Varlet MN, Sung RT. Toxoplasma gondii serology in pregnant woman: characteristics and pitfalls. Ann Biol Clin (Paris) 2009; 67: 125-133.
  3. Meroni V, Genco F. Toxoplasmosis in pregnancy: evaluation of diagnostic methods. Parassitologia 2008; 50: 51-53.
  4. Sensini A. Toxoplasma gondii infection in pregnancy: opportunities and pitfalls of serological diagnosis. Clin Microbiol Infect 2006; 12: 504-512. https://doi.org/10.1111/j.1469-0691.2006.01444.x
  5. Khammari I, Saghrouni F, Yaacoub A, Gaied Meksi S, Ach H, Garma L, Fathallah A, Ben Saïd M. IgG western blot for confirmatory diagnosis of equivocal cases of toxoplasmosis by EIA-IgG and fluorescent antibody test. Korean J Parasitol 2013; 51: 485-488. https://doi.org/10.3347/kjp.2013.51.4.485
  6. Franck J, Garin YJ, Dumon H. LDBio-Toxo II immunoglobulin G Western blot confirmatory test for anti-toxoplasma antibody detection. J Clin Microbiol 2008; 46: 2334-2338. https://doi.org/10.1128/JCM.00182-08
  7. Cimon B, Marty P, Morin O, Bessieres MH, Marx-Chemla C, Gay-Andrieu F, Thulliez P. Specificity of low anti-Toxoplasma IgG titers with IMx and AxSYM Toxo IgG assays. Diagn Microbiol Infect Dis 1998; 32: 65-67. https://doi.org/10.1016/S0732-8893(98)00060-1
  8. Derouin F, Garin YJ, Buffard C, Berthelot F, Petithory JC. Multicenter study of toxoplasma serology using various commercial ELISA reagents. Work Group Toxoplasmosis of the National Agency for Quality Control in Parasitology. Bull WHO 1994; 72: 249-256.
  9. Petithory JC, Ambroise-Thomas P, De Loye J, Pelloux H, Goullier-Fleuret A, Milgram M, Buffard C, Garin JP. Serodiagnosis of toxoplasmosis: a comparative multicenter study of a standard scale through various actual tests and expression of the results in international units. Bull WHO 1996; 74: 291-298.
  10. Lesle F, Touafek F, Fekkar A, Mazier D, Paris L. Discrepancies between a new highly sensitive Toxoplasma gondii ELISA assay and other reagents: interest of Toxo IgG western blot. Eur J Clin Microbiol Infect Dis 2011; 30: 1207-1212. https://doi.org/10.1007/s10096-011-1214-1
  11. Kahi S, Cozon GJ, Greenland T, Wallon M, Gay-Andrieu F, Peyron F. A rapid flow cytometric method to explore cellular immunity against Toxoplasma gondii in humans. Clin Diagn Lab Immunol 1998; 5:745-748.
  12. Ma GY, Zhang JZ, Yin GR, Zhang JH, Meng XL, Zhao F. Toxoplasma gondii: proteomic analysis of antigenicity of soluble tachyzoite antigen. Exp Parasitol 2009; 122: 41-46. https://doi.org/10.1016/j.exppara.2009.01.011
  13. Laemmli 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
  14. Perkins NJ, Schisterman EF. The Youden Index and the optimal cut-point corrected for measurement error. Biom J 2005; 47: 428-441. https://doi.org/10.1002/bimj.200410133
  15. Reiter-Owona I, Petersen E, Joynson D, Aspock H, Darde ML, Disko R, Dreazen O, Dumon H, Grillo R, Gross U, Hayde M, Holliman R, Ho-Yen DO, Janitschke K, Jenum PA, Naser K, Olszewski M, Thulliez P, Seitz HM. The past and present role of the Sabin-Feldman dye test in the serodiagnosis of toxoplasmosis. Bull WHO 1999; 77: 929-935.
  16. Gendrin C, Bittame A, Mercier C, Cesbron-Delauw MF. Post-translational membrane sorting of the Toxoplasma gondii GRA6 protein into the parasite-containing vacuole is driven by its N-terminal domain. Int J Parasitol 2010; 40: 1325-1334. https://doi.org/10.1016/j.ijpara.2010.03.014
  17. Lovgren K, Uggla A, Morein B. A new approach to the preparation of a Toxoplasma gondii membrane antigen for use in ELISA. Zentralbl Veterinarmed B 1987; 34: 274-282.
  18. Sharma SD, Mullenax J, Araujo FG, Erlich HA, Remington JS. Western blot analysis of the antigens of Toxoplasma gondii recognized by human IgM and IgG antibodies. J Immunol 1983; 131: 977-983.
  19. Marcolino PT, Silva DA, Leser PG, Camargo ME, Mineo JR. Molecular markers in acute and chronic phases of human toxoplasmosis: determination of immunoglobulin G avidity by western blotting. Clin Diagn Lab Immunol 2000; 7: 384-389.
  20. Reichmann G, Dlugonska H, Fischer HG. Characterization of TgROP9 (p36), a novel rhoptry protein of Toxoplasma gondii tachyzoites identified by T cell clone. Mol Biochem Parasitol 2002; 119: 43-54. https://doi.org/10.1016/S0166-6851(01)00397-8
  21. Fatoohi AF, Cozon GJ, Gonzalo P, Mayencon M, Greenland T, Picot S, Peyron F. Heterogeneity in cellular and humoral immune responses against Toxoplasma gondii antigen in humans. Clin Exp Immunol 2004; 136: 535-541. https://doi.org/10.1111/j.1365-2249.2004.02466.x
  22. Attallah AM, Ismail H, Ibrahim AS, Al-Zawawy LA, El-Ebiary MT, El-Waseef AM. Immunochemical identification and detection of a 36-kDa Toxoplasma gondii circulating antigen in sera of infected women for laboratory diagnosis of toxoplasmosis. J Immunoassay Immunochem 2006; 27: 45-60. https://doi.org/10.1080/15321810500403748
  23. Johnson AM, McDonald PJ, Neoh SH. Molecular weight analysis of soluble antigens from Toxoplasma gondii. J Parasitol 1983; 69: 459-464. https://doi.org/10.2307/3281354
  24. Holec-Gasior L, Kur J. Toxoplasma gondii: recombinant GRA5 antigen for detection of immunoglobulin G antibodies using enzyme-linked immunosorbent assay. Exp Parasitol 2010; 124: 272-278. https://doi.org/10.1016/j.exppara.2009.10.010
  25. Ali-Heydari S, Keshavarz H, Shojaee S, Mohebali M. Diagnosis of antigenic markers of acute toxoplasmosis by IgG avidity immunoblotting. Parasite 2013; 20: 18. https://doi.org/10.1051/parasite/2013017

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