Obtaining an ELISA test based on a recombinant protein of Chlamydia trachomatis

  • María J. de Haro-Cruz
  • Sandra I. Guadarrama-Macedo
  • Marcela López-Hurtado
  • Marcos R. Escobedo-Guerra
  • Fernando M. Guerra-InfanteEmail author
Original Article


Chlamydia trachomatis is considered as a public health problem due to its high prevalence and increased rates of gynecological disorders. The major outer membrane protein (MOMP) of this bacterium is the most abundant protein in its membrane and has been evaluated not only as a vaccine development candidate but also is used in many diagnostic tests. The MOMP weighs 69 kDa and contains four variable segments (VS 1–4) separated by constant regions. Several research groups have developed recombinant single-variable segments of MOMP expressed in Escherichia coli cytoplasm. But, all variable segments have been used minimally for the diagnosis of a chlamydial infection. In this experiment, the authors obtained the recombinant MOMP of C. trachomatis (rMOMP) in E. coli rMOMP and extracted, purified, and partially characterized it. This was later used to identify anti-Chlamydia trachomatis antibodies in sera of infertile patients by immunodetection assays, enzyme-linked immunosorbent assay (ELISA), and indirect immunofluorescence tests. The ELISA test showed high sensitivity and low specificity of 100 and 58.3%, respectively. The above results obtained were linked to the cross-reactivity of antibodies against C. pneumoniae or C. psittaci. Hence, an evaluation was performed to obtain an optimized test for the diagnosis of C. trachomatis infection.


Chlamydia trachomatis MOMP of Chlamydia Recombinant proteins Chlamydia diagnosis 


Authors’ contribution

The authors have seen and approved the manuscript being submitted.

Compliance with ethical standards

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Furthermore, Informed consent was obtained from all individual participants involved in the study.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Bandehpour M, Seyed N, Shadnoush M, Pakzad P, Kazemi B (2006) Using recombinant Chlamydia major outer membrane protein (MOMP) in ELISA diagnostic Kit. Iran J Biotechnol 4:239–244. RetrievedMarch 26, 2019,
  2. Bastidas RJ, Elwell CA, Engel JN, Valdivia RH (2013) Chlamydial intracellular survival strategies. Cold Spring Harb Perspect Med 3:a010256. CrossRefGoogle Scholar
  3. Batteiger BE, Lin PM, Jones RB, Van Der Pol BJ (1996) Species-, serogroup-, and serovar-specific epitopes are juxtaposed in variable sequence region 4 of the major outer membrane proteins of some Chlamydia trachomatis serovars. Infect Immun 64:2839–2841Google Scholar
  4. Baud D, Regan L, Greub G (2010) Comparison of five commercial serological tests for the detection of anti-Chlamydia trachomatis antibodies. Eur J Clin Microbiol Infect Dis 29:669–675. CrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  6. Cui J, Yan W, Xie H, Xu S, Wang Q, Zhang W, Ni A (2018) A retrospective seroepidemiologic survey of Chlamydia pneumoniae infection in patients in Beijing between 2008 and 2017. PLoS ONE 13:e0206995.
  7. De Haro-Cruz M, DeLeón-Rodriguez I, Escobedo-Guerra MR, López-Hurtado M, Arteaga-Troncoso G, Ortiz-Ibarra FJ, Guerra-Infante FM (2011) Genotyping of Chlamydia trachomatis from endocervical specimens of infertile Mexican women. Enferm Infecc Microbiol Clin 29:102–108. CrossRefGoogle Scholar
  8. Fenga C, Cacciola A, Di Nola C, Calimeri S, Lo Giudice D, Pugliese M, Niutta PP, Martino LB (2007) Serologic investigation of the prevalence of Chlamydophila psittaci in occupationally-exposed subjects in eastern Sicily. Ann Agric Environ Med 14:93–96Google Scholar
  9. Findlay HE, McClafferty H, Ashley RH (2005) Surface expression, single-channel analysis and membrane topology of recombinant Chlamydia trachomatis major outer membrane protein. BMC Microbiol 26:5. CrossRefGoogle Scholar
  10. Hernández-Trejo M, Herrera-González N, Guerra-Infante FM (2014) Serological evidence of infection by three species of Chlamydia in pregnant women. Ginecol Obstet Mex 82:585–590Google Scholar
  11. Ishizaki M, Allen JE, Beatty PR, Stephens RS (1992) Immune specificity of murine T-cell lines to the major outer membrane protein of Chlamydia trachomatis. Infect Immun 60:3714–3718Google Scholar
  12. Jiang P, Cai Y, Chen J, Ye X, Mao S, Zhu S, Xue X, Chen S, Zhang L (2017) Evaluation of tandem Chlamydia trachomatis MOMP multi-epitopes vaccine in BALB/c mice model. Vaccine 35:3096–3103.
  13. Jones CS, Maple PA, Andrews NJ, Paul ID, Caul EO (2003) Measurement of IgG antibodies to Chlamydia trachomatis by commercial enzyme immunoassays and immunofluorescence in sera from pregnant women and patients with infertility, pelvic inflammatory disease, ectopic pregnancy, and laboratory diagnosed Chlamydia psittaci/Chlamydia pneumoniae infection. J Clin Pathol 56:225–229CrossRefGoogle Scholar
  14. Khan S, Ullah MW, Siddique R, Nabi G, Manan S, Yousaf M, Hou H (2016) Role of recombinant DNA technology to improve Life. Int J Genomics 2016:2405954. Google Scholar
  15. Land JA, Gijsen AP, Kessels AG, Slobbe ME, Bruggeman CA (2003) Performance of five serological chlamydia antibody tests in subfertile women. Hum Reprod 18:2621–2627CrossRefGoogle Scholar
  16. Menon S, Stansfield SH, Walsh M, Hope E, Isaia L, Righarts AA, Niupulusu T, Temese SV, Iosefa-Siitia L, Auvaa L, Tapelu SA, Motu MF, Suaalii-Sauni T, Timms P, Hill PC, Huston WM (2016) Sero-epidemiological assessment of Chlamydia trachomatis infection and sub-fertility in Samoan women. BMC Infect Dis 16:175. CrossRefGoogle Scholar
  17. Moss TR, Darougar S, Woodland RM, Nathan M, Dines RJ, Cathrine V (1993) Antibodies to Chlamydia species in patients attending a genitourinary clinic and the impact of antibodies to C. pneumoniae and C. psittaci on the sensitivity and the specificity of C. trachomatis serology tests. Sex Transm Dis 20:61–65CrossRefGoogle Scholar
  18. Mygind P, Christiansen G, Persson K, Birkelund S (2000) Detection of Chlamydia trachomatis-specific antibodies in human sera by recombinant major outer-membrane protein polyantigens. J Med Microbiol 49:457–465CrossRefGoogle Scholar
  19. O’Connell CM, Ferone ME (2016) Chlamydia trachomatis genital infections. Microb Cell 3:390–403. CrossRefGoogle Scholar
  20. Rahman KS, Darville T, Russell AN, O’Connell CM, Wiesenfeld HC, Hillier SL, Chowdhury EU, Juan YC, Kaltenboeck B (2018a) Discovery of human-specific immunodominant Chlamydia trachomatis B cell epitopes. mSphere. 3:e00246–e00218. Google Scholar
  21. Rahman KS, Darville T, Russell AN, O’Connell CM, Wiesenfeld HC, Hillier SL, Lee DE, Kaltenboeck B (2018b) Comprehensive molecular serology of human Chlamydia trachomatis infections by peptide enzyme-linked immunosorbent assays. mSphere 3:e00253–e00218. Google Scholar
  22. Sardiu ME, Cheung MS, Yu YK (2007) Cysteine-cysteine contact preference leads to target-focusing in protein folding. Biophys J 93:938–951CrossRefGoogle Scholar
  23. Schumann W, Ferreira LCS (2004) Production of recombinant proteins in Escherichia coli. Genet Mol Biol 27:442–453CrossRefGoogle Scholar
  24. Sun G, Pal S, Sarcon AK, Kim S, Sugawara E, Nikaido H, Cocco MJ, Peterson EM, de la Maza LM (2007) Structural and functional analyses of the major outer membrane protein of Chlamydia trachomatis. J Bacteriol 189:6222–6235CrossRefGoogle Scholar
  25. Wen Z, Boddicker MA, Kaufhold RM, Khandelwal P, Durr E, Qiu P, Lucas BJ, Nahas DD, Cook JC, Touch S, Skinner JM, Espeseth AS, Przysiecki CT, Zhang L (2016) Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. coli outer membrane as a substrate for vaccine research. BMC Microbiol 16:165. CrossRefGoogle Scholar
  26. Yang CL, Maclean I, Burnham RC (1993) DNA sequence polymorphism of the Chlamydia trachomatis omp1 gene. J Infect Dis 168:1225–1230CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Department of Microbiology, Escuela Nacional de Ciencias Biológicas del Instituto Politécnico NacionalMexico CityMéxico
  2. 2.Virology Laboratory, National Institute of Perinatology, Isidro Espinoza de los ReyesMexico CityMexico

Personalised recommendations