Ukr.Biochem.J. 2017; Volume 89, Issue 1, Jan-Feb, pp. 22-30


Characteristics of enzyme-linked immunosorbent assay for detection of IgG antibodies specific to Сhlamydia trachomatis heat shock protein (HSP-60)

O. Yu. Galkin, A. B. Besarab, T. N. Lutsenko

National Technical University of Ukraine “Igor Sikorsky Kyiv Polytechnic Institute”;

The goal of this work was to study sensitivity and specificity of the developed ELISA set for the identification of IgG antibodies against Chlamydia trachomatis HSP-60 (using biotinylated tyramine-based signal amplification system). The study was conducted using a panel of characterized sera, as well as two reference ELISA sets of similar purpose. According to the results of ELISA informative value parameters, the ELISA we have developed showed the highest specificity and sensitivity parameters (no false negative or false positive results were registered). In 4 out of 15 intralaboratory panel serum samples initially identified as negative, anti-HSP-60 IgG-antibodies test result in reference ELISA sets upon dilution changed from negative to positive. The nature of titration curves of false negative sera and commercial monoclonal antibodies А57-В9 against C. trachomatis HSP-60 after incubation for 24 h was indicative of the presence of anti-idiotypic antibodies in these samples. Upon sera dilution, idiotypic-anti-idiotypic complexes dissociated, which caused the change of test result. High informative value of the developed ELISA set for identification of IgG antibodies against C. trachomatis HSP-60 has been proven. Anti-idiotypic antibodies possessing C. trachomatis anti-HSP-60 activity and being one of the causes of false negative results of the relevant ELISA-based tests have been identified in blood sera of individuals infected with chlamydial genitourinary infection agents.

Keywords: , , , , ,


  1. Mavrov GI, Scherbakova YuV, Chinov GP. The treatment of complicated chamydial infection with azithromycin (Sumamed) using in combination with pathogenetic therapy. Ukr J Dermatol Venerol Cosmetol. 2010; (3(38)): 123-127. (In Russian).
  2. Medical microbiology, virology, immunology. Ed. V.P. Shyrobokov. Vinnitsa: Nova knyga, 2011. 951 p. (In Ukrainian).
  3. Biotechnological bases of creation of medical devises for serological diagnosis of infectious and non-infectious diseases: Monograph / O.Yu. Galkin, V.P. Shyrobokov, A.A. Grigorenko, O.M. Dugan, T.M. Lutsenko, A.G. Komar; Ed. V.P. Shyrobokov. Kyiv: NTUU “KPI”, 2015. 204 p. (In Ukrainian).
  4. Galkin OYu, Besarab OB, Gryshyna AS, Dugan OM, Gurzhenko YuM. Biotechnology for obtaining the recombinant heat shock protein (HSP-60) of Chlamydia trachomatis and evaluation of the perspectives of its use in serological diagnostics. Sci Herald Chernivtsi  Univ. Biology (Biol System). 2014; 6(2): 130-138.
  5. Rakhmatullina MR, Popov DV, Plakhova KI. Current concepts of the epidemiology, clinical picture, diagnostics and therapy of non-complicated and complicated forms of the urogenital chlamydia infection in men. Vestnik Dermatol Venerol. 2012; 6: 35-41. (In Russian).
  6. Galkin ОYu, Solovjova VF, Metalnikova NP, Dugan ОM, Gorgo YuP. Obtaining of monoclonal antibodies to human IgG suitable for usage in highly sensitive and specific immunoassays. Visnyk Zaporizh Nat Univ. Biol Sci. 2014; 2: 120-129.
  7. Jensen IP, Fogh H, Prag J. Diagnosis of Chlamydia trachomatis infections in a sexually transmitted disease clinic: evaluation of a urine sample tested by enzyme immunoassay and polymerase chain reaction in comparison with a cervical and/or a urethral swab tested by culture and polymerase chain reaction. Clin Microbiol Infect. 2003 Mar;9(3):194-201. PubMed, CrossRef
  8. Bébéar C, de Barbeyrac B. Genital Chlamydia trachomatis infections. Clin Microbiol Infect. 2009 Jan;15(1):4-10. Review. PubMed, CrossRef
  9. Baud D, Zufferey J, Hohlfeld P, Greub G. Performance of an automated multiplex immunofluorescence assay for detection of Chlamydia trachomatis immunoglobulin G. Diagn Microbiol Infect Dis. 2014 Mar;78(3):217-9. PubMed, CrossRef
  10. Sulaiman S, Chong PP, Mokhtarudin R, Lye MS, Wan Hassan WH. Comparison of nested and ELISA based polymerase chain reaction assays for detecting Chlamydia trachomatis in pregnant women with preterm complications. Trop Biomed. 2014 Mar;31(1):36-45. PubMed
  11. Isakov VA, Kulyashova LB, Berezina LA, Svarval AV. Laboratory diagnosis of urogenital chlamydiosis. 2. Modern methods of diagnostics of chlamydia infections: analytical review. Terra Medica Labor Diagn. 2013; 1: 13-21. (In Russian).
  12. Galkin OYu, Kazmirchuk VYe, Metal’nikova NP. Heat Shock Proteins: Role in Formation of Immune Response. Res Bull NTUU “KPI”.  2014; 3: 12-20. (In Ukrainian).
  13. Giuliano JS Jr, Lahni PM, Wong HR, Wheeler DS. Pediatric Sepsis – Part V: Extracellular Heat Shock Proteins: Alarmins for the Host Immune System. Open Inflamm J. 2011 Oct 7;4:49-60. PubMed, PubMedCentral, CrossRef
  14. Pockley AG. Heat shock proteins as regulators of the immune response. Lancet. 2003 Aug 9;362(9382):469-76. PubMed, CrossRef
  15. Habich C, Kempe K, Gomez FJ, Lillicrap M, Gaston H, van der Zee R, Kolb H, Burkart V. Heat shock protein 60: identification of specific epitopes for binding to primary macrophages. FEBS Lett. 2006 Jan 9;580(1):115-20. PubMed, CrossRef
  16. Behn U. Idiotypic networks: toward a renaissance? Immunol Rev. 2007 Apr;216:142-52. Review. PubMed, CrossRef
  17. Bobrovnik SA. Ligand-receptor interactions: a new method for determining the binding parameters. J Biochem Biophys Methods. 2003 Jan 31;55(1):71-86. PubMed, CrossRef
  18. Bobrovnik SA. Idiotype-antiidiotypic interaction of antibodies in the bloodstream. Ukr Biochem J. 2014 Mar-Apr;86(2):89-100. (In Russian). PubMed, CrossRef
  19. Stefatić D, Riederer M, Balić M, Dandachi N, Stanzer S, Janesch B, Resel M, Ler D, Samonigg H, Bauernhofer T. Optimization of diagnostic ELISA-based tests for the detection of auto-antibodies against tumor antigens in human serum. Bosn J Basic Med Sci. 2008 Aug;8(3):245-50. PubMed
  20. Ivanskaya NV. Significance of antigenic mimicry when determining antibodies to human immunodeficiency virus in sera of patients with tuberculosis. Lab Diagn. 2011; (1(55)): 39-44.

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