ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)
Biotechnologia Acta V. 18, No. 6, 2025
P. 13-25, Bibliography 68, Engl.
UDC: 602+579+616.9
doi: https://doi.org/10.15407/biotech18.06.013
PROBLEMS AND STRATEGIES IN THE DETECTION OF Mycoplasma hominis
L.V. VLASENKO, T.S. TODOSIICHUK
National Technical University of Ukraine “Igor Sikorsky Kyiv Polytechnic Institute”
Among the diseases that are transmitted mainly sexually, urogenital mycoplasmosis occupies one of the first places. Clinical manifestations of the disease caused by M. hominis are often similar to the symptoms of other diseases of the urogenital tract of bacterial, viral and other etiologies. Therefore, the diagnosis of urogenital mycoplasmosis caused by M. hominis faces a number of problems, despite the possibility of using genetic methods for direct determination of the DNA of this microorganism.
Aim. Comprehensive analysis of the features of M. hominis and modern methods to determine promising approaches in the diagnosis of urogenital mycoplasmosis caused by this microorganism.
Methods. Search and systematization of modern scientific data and results on the features of the detection of mycoplasmas in the literature, covering numerous databases, including PubMed, Web of Science, Scopus, Google Scholar and other relevant sources.
Results. A comprehensive analysis of the features of the morphology, genetics and mechanisms of interaction with the human body of M. hominis was carried out. The possibilities of modern methods for detecting this pathogen of urogenital mycoplasmosis are considered, including genetic, immunological,
immunocytochemical, immunohistochemical methods, as well as immunochromatographic tests, lateral flow assays or rapid tests.
Conclusions. The complexity of detecting M. hominis in the process of diagnosing urogenital mycoplasmosis is due to the morphological and genetic features of the pathogen, various mechanisms of masking from the immune system, its presence in the human body normally and in combination with other pathogens. The most effective strategy for detecting mycoplasmas in the diagnosis of diseases is the simultaneous determination of their antigens in urogenital samples and antibodies in the blood, which can also be a guideline for improving or developing new complex test systems.
Keywords: M. hominis, urogenital mycoplasmosis, polymerase chain reaction, enzyme-linked immunosorbent assay, antigens, antibodies, diagnostics.
© Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 2025
References
1. Dessì, D., Margarita, V., Cocco, A. R., Marongiu, A., Fiori, P. L., Rappelli, P. (2019). Trichomonas vaginalis and Mycoplasma hominis: new tales of two old friends. Parasitology, (146)9, 1150–1155. https://doi.org/10.1017/S0031182018002135
2. Horner, P., Donders, G., Cusini, M., Gomberg, M., Jensen, J. S., Unemo, M. (2018). Should we be testing for urogenital Mycoplasma hominis, Ureaplasma parvum and Ureaplasma urealyticum in men and women? – a position statement from the European STI Guidelines Editorial Board. J Eur Acad Dermatol Venereol, (32)11, 1845–1851. https://doi.org/10.1111/jdv.15146
3. Jensen, J. S., Cusini, M., Gomberg, M., Moi, H., Wilson, J., Unemo, M. (2022). 2021 European guideline on the management of Mycoplasma genitalium infections. J Eur Acad Dermatol Venereol, (36)5, 641–650. https://doi.org/10.1111/jdv.17972
4. Baumann, L., Cina, M., Egli-Gany, D., Goutaki, M., Halbeisen, F.S., Lohrer, G.-R., Ali, H., Scott, P., ..., Low, N. (2018). Prevalence of Mycoplasma genitalium in different population groups: systematic review and meta-analysis. Sex Transm Infect, (94)4, 255–262. https://doi.org/10.1136/sextrans-2017-053384
5. Gu, X., Liu, S., Guo, X., Weng, R., Zhong, Z. (2020). Epidemiological investigation and antimicrobial susceptibility analysis of mycoplasma in patients with genital manifestations. J Clin Lab Anal, (34)4, 1–6. https://doi.org/10.1002/jcla.23118
6. Cai, S., Pan, J., Wang, S., Bao, R., Zhou, C., Gao X., Shiiya H., Hu B. (2024). Accurate etiological diagnosis of Mycoplasma hominis mediastinitis in immunocompetent patients using metagenomic next-generation sequencing: a case series and literature review. J. Thorac. Dis, (16)4, 2499–2509. https://doi.org/10.21037/jtd-24-286
7. Carneiro, F. P., Darós, A. C., Darós, A. C. M., Mendes Lousa de Castro, T. M., Carneiro, M. V., Fidelis, C. R., Vilioni, M. V., Matsunaga, M. E. da C., ..., Motoyama, A. B. (2020). Cervical Cervical Cytology of Samples with Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Trichomonas vaginalis, Mycoplasma hominis, and Neisseria gonorrhoeae Detected by Multiplex PCR. BioMed Res. Int, (2020)1, 1–10. https://doi.org/10.1155/2020/7045217
8. Tamargo M.I. (2016). Culture medium for the isolation of clinical samples such Mycoplasma including which are not culturable in traditional media. (Italy. Pat. No. WO 2016/157109). https://www.wipo.int/portal/en/
9. Yiwen, C., Yueyue, W., Lianmei, Q., Cuiming, Z., Xiaoxing, Y. (2021). Infection strategies of mycoplasmas: Unraveling the panoply of virulence factors. Virulence, (12)1, 788–817. https://doi.org/10.1080/21505594.2021.1889813
10. Rideau, F., Le Roy, C., Sagné, E., Renaudin, H., Pereyre, S., Henrich, B. Dordet-Frisoni, E., Citti, C., ..., Bébéar, C. (2019). Random transposon insertion in the Mycoplasma hominis minimal genome. Sci Rep, (9)1, 13554. https://doi.org/10.1038/s41598-019-49919-y
11. Pereyre, S., Sirand-Pugnet, P., Beven, L., Charron, A., Renaudin, H., Barré, A., Avenaud, P., Jacob, D. ..., Bébéar, C. (2009). Life on Arginine for Mycoplasma hominis: Clues from Its Minimal Genome and Comparison with Other Human Urogenital Mycoplasmas. PLOS Genet, (5)10, e1000677. https://doi.org/10.1371/journal.pgen.1000677
12. Boujemaa, S., Ben Allaya, A., Mlik, B., Mardassi, H., Ben Abdelmoumen Mardassi, B. (2018). Phylogenetics of Mycoplasma hominis clinical strains associated with gynecological infections or infertility as disclosed by an expanded multilocus sequence typing scheme. Sci Rep, (8), 14854. https://doi.org/10.1038/s41598-018-33260-x
13. Xiu, F., Li, X., Liu, L., Xi, Y., Yi, X., Li, Y. You, X. (2024). Mycoplasma invasion into host cells: An integrated model of infection strategy. Molecular Microbiology, (121)4, 814–830. https://doi.org/10.1111/mmi.15232
14. Benedetti, F., Curreli, S., Zella, D. (2020). Mycoplasmas–Host Interaction: Mechanisms of Inflammation and Association with Cellular Transformation. Microorganisms, (8)9, 1351. https://doi.org/10.3390/microorganisms8091351
15. Al-Ezzi, N. J., & Al-Thahab, A. (2022). Studying the relationship between tlr-6 and mycoplasmas among urinary-genital tract infections for females in al-hillah city, Iraq. International Journal of Health Sciences, (6)S7, 905–914. https://doi.org/10.53730/ijhs.v6nS7.11498
16. Henrich, B., Feldmann, R. C., Hadding, U. (1993). Cytoadhesins of Mycoplasma hominis. Infection and immunity, (61)7, 2945–2951. https://doi.org/10.1128/iai.61.7.2945-2951.1993
17. Brown, R. J., Chalker, V. J., Spiller, O. B. (2014). Mycoplasma hominis Variable Adherence-Associated Antigen: A Major Adhesin and Highly Variable Surface Membrane Protein. Advances in Microbiology, (4), 736–746. https://doi.org/10.4236/aim.2014.411080
18. Saadat, S., Sajadi, M. M., Alikhani, M. Y., Rikhtegaran Tehrani, Z. Yousefi Mashouf, R. (2018). Production of a chimeric protein and its potential application in sero-diagnosis of Mycoplasma hominis infection. Journal of Microbiological Methods, (144), 186–191. https://doi.org/10.1016/j.mimet.2017.12.001
19. Wang, J., Liang, K., Chen, L., Su, X., Liao, D., Yu, J., He, J. (2023). Unveiling the stealthy tactics: mycoplasma’s immune evasion strategies. Front. Cell. Infect. Microbiol, (13). https://doi.org/10.3389/fcimb.2023.1247182
20. Christiansen, G., Søren, L.M, Nyvold, C., Birkelund, S. (1994). Analysis of a Mycoplasma hominis membrane protein, P120. FEMS Microbiology Letters, (121)1, 121–127. https://doi.org/10.1111/j.1574-6968.1994.tb07085.x
21. Ben Abdelmoumen Mardassi, B., Ayari, H., Béjaoui-Khiari, A., Mlik, B., Moalla, I., Amouna, F. (2007). Genetic variability of the P120′ surface protein gene of Mycoplasma hominis isolates recovered from Tunisian patients with uro-genital and infertility disorders. BMC Infect Dis, (7), 142. https://doi.org/10.1186/1471-2334-7-142
22. Amorim, A. T., Lino, V. de S., Marques, L. M., Martins, D. J., Braga Junior, A. C. R., Campos, G. B., Oliveira, C. N. T., Boccardo, E., ..., Timenetsky, J. (2022). Mycoplasma hominis Causes DNA Damage and Cell Death in Primary Human Keratinocytes. Microorganisms, (10)10, 1962. https://doi.org/10.3390/microorganisms10101962
23. Hopfe, M., Dahlmanns, T. Henrich, B. (2011). In Mycoplasma hominis the OppA-mediated cytoadhesion depends on its ATPase activity. BMC Microbiol, 11, (185)2011. https://doi.org/10.1186/1471-2180-11-185
24. Kitzerow, A., Hadding, U., Henrich, B. (1999). Cyto-adherence studies of the adhesin P50 of Mycoplasma hominis. Journal of Medical Microbiology. (48)5, 485–493. https://doi.org/10.1099/00222615-48-5-485
25. Zakariah, M., Khan, S., Chaudhary, A. A., Rolfo, C., Ben Ismail, M. M., Alotaibi, Y. A. (2018). To decipher the Mycoplasma hominis proteins targeting into the endoplasmic reticulum and their implications in prostate cancer etiology using Next-Generation Sequencing data. Molecules, (23)5, 994. https://doi.org/10.3390/molecules23050994
26. Khan, S., Zakariah, M., Rolfo, C., Robrecht, L., Palaniappan, S. (2017). Prediction of mycoplasma hominis proteins targeting in mitochondria and cytoplasm of host cells and their implication in prostate cancer etiology. Oncotarget, (8), 30830–30843. https://doi.org/10.18632/oncotarget.8306
27. Arfi, Y., Lartigue, C., Sirand-Pugnet, P., Blanchard, A. (2021). Beware of Mycoplasma anti-immunoglobulin strategies. MBio, (12)6, e01974-21. https://doi.org/10.1128/mBio.01974-21
28. Shiragannavar, S., Madagi, S. (2020). In silico vaccine design against mycoplasma hominis infections. Biomed. Pharmacol. J, (13)1, 457–468. https://dx.doi.org/10.13005/bpj/1906
29. Margarita, V., Cao, L. C., Bailey, N. P., Ngoc, T. H. T., Ngo, T. M. C., Nu, P. A. T., Diaz, N., Dessì, D., …, Rappelli, P. (2022). Effect of the Symbiosis with Mycoplasma hominis and Candidatus Mycoplasma Girerdii on Trichomonas vaginalis Metronidazole Susceptibility. Antibiotics, (11)6, 812. https://doi.org/10.3390/antibiotics11060812
30. Rudenko, A.V., Mitchenko, M.V., Bavina, O.M., Tretiak, V.V. (2019). Comparative characteristics of the culture-enzymatic and molecular-genetic methods for indicating mollicutes – Mycoplasma hominis and Ureaplasma spp., isolated in patients with acute uncomplicated pyelonephritis. Health of man, (4), 52–56. (in Ukrainian). https://doi.org/10.30841/2307-5090.4.2019.195269
31. ELITechGroup. MYCOFAST®REVOLUTION. Urogenital mycoplasma diagnosis. Direct method. https://www.elitechgroup.com/documentation/wp-content/uploads/sites/30/2022/12/Brochure_MYCOFAST-Rev2-Direct_EN_2025.pdf
32. Dósa, E., Nagy, E., Falk, W., Szőke, I., Ballies, U. (1999). Evaluation of the Etest for susceptibility testing of Mycoplasma hominis and Ureaplasma urealyticum. J. Antimicrob. Chemother, (43)4, 575–578. https://doi.org/10.1093/jac/43.4.575
33. D’Inzeo, T., De Angelis, G., Fiori, B., Menchinelli, G., Liotti, FM., Morandotti, GA., De Maio, F., Nagel, D., …, Spanu, T. (2017). Comparison of Mycoplasma IES, Mycofast Revolution and Mycoplasma IST2 to detect genital mycoplasmas in clinical samples. J Infect Dev Ctries, (11)01, 98–101. https://doi.org/10.3855/jidc.8039
34. Morris, D.J., Jones, L.C., Davies, R.L. Sands, K., Portal E., Spiller, O. B.(2020). MYCO WELL D-ONE detection of Ureaplasma spp. and Mycoplasma hominis in sexual health patients in Wales. Eur. J. Clin. Microbiol. Infect. Dis, (39)12, 2427–2440. https://doi.org/10.1007/s10096-020-03993-7
35. Saavedra-Alonso, S., Chávez-Escamilla, A. K., Escobedo-Guajardo, B. L., Alvarez-Ojeda, G. González-Villarreal, C. A., Moreno-Treviño, M. G., Lugo-Ibarra, J. F. (2025). Advancements in Molecular Diagnostics for Viral Outbreaks: A Focus on Respiratory Virus. IntechOpen. https://doi.org/10.5772/intechopen.1011431
36. Aycan, M., Yildiz, M. (2024). Introductory Chapter: A Molecular Photocopying Polymerase Chain Reaction (PCR). IntechOpen. https://doi.org/10.5772/intechopen.113879
37. Johnson, J.S., Spakowicz, D.J., Hong, B.Y., Petersen, L.M., Demkowicz, P., Chen, L., Leopold, S.R., Hanson, B.M., ..., Weinstock, G.M. (2019). Evaluation of 16S rRNA gene sequencing for species and strain-level microbiome analysis. Nature Communications, (10)5029. https://doi.org/10.1038/s41467-019-13036-1
38. Xiu, L, Zhang, C, Li, Y, Wang, F, Peng, J. (2019). Simultaneous detection of eleven sexually transmitted agents using multiplexed PCR coupled with MALDI-TOF analysis. Infect. Drug Resist, (12)6, 2671–2682. https://doi.org/10.2147/IDR.S219580
39. Lesiak-Markowicz, I., Walochnik, J., Stary, A., Fürnkranz, U. (2023). Detection of Putative Virulence Genes alr, goiB, and goiC in Mycoplasma hominis Isolates from Austrian Patients. International Journal of Molecular Sciences, (24)9, 7993. https://doi.org/10.3390/ijms24097993
40. Moridi, K., Ghazvini, K., Hemmati, M., Hoseiniun, H., Torkaman, M., Fallah Mehrabadi, M. H. (2021). Prevalence Determination of M. Hominis and M. Genitalium in the Semen Samples in the Northeast of Iran Using Culture and Multiplex Polymerase Chain Reaction. Archives of Razi Institute, (76)1, 41–49. https://doi.org/10.22092/ari.2019.125966.1338
41. Férandon, C., Peuchant, O., Janis, C., Benard, A., Renaudin, H., Pereyre, S., Bébéar, C. (2011). Development of a real-time PCR targeting the yidC gene for the detection of Mycoplasma hominis and comparison with quantitative culture. Clinical Microbiology and Infection, (17)2, 155–159. https://doi.org/10.1111/j.1469-0691.2010.03217.x
42. NZYtech. Mycoplasma hominis qPCR Kit. Instructions for use. https://www.nzytech.com/media/dds/brochurescertificates/md0763_ifu_en_v2402.pdf
43. Razin S., Yogev D., Naot Y. (1998). Molecular Biology and Pathogenicity of Mycoplasmas. Microbiol. Mol. Biol. Rev., (62)4, 1094–1156. https://doi.org/10.1128/MMBR.62.4.1094-1156.1998
44. GeneProof. GeneProof Mycoplasma genitalium/hominis PCR Kit. https://static.viaaurea.eu/d/geneproofcom/files/12296.pdf/s-42c2a65b5da0?fn=GP_MGH_Leaflet_EN.pdf
45. Elisabeth Pharmacon. Sexually Transmitted Urinary Infection. EliGene®. Mycoplasma hom/gen UNI. https://www.elisabeth.cz/en/product-mycoplasma-hom-gen-uni-en.html
46. Soheily, Z., Soleimani, M., Majidzadeh-Ardebili, K. (2018). Detection of Mycoplasma Contamination of Cell Culture by A Loop-Mediated Isothermal Amplification Method. Cell Journal (Yakhteh), (21)1, 43–48. https://doi.org/10.22074/cellj.2019.5624
47. InvivoGen. MycoStripTM 100. https://www.invivogen.com/mycostrip
48. Instruction Manual. Research Use Only. MPbio. Myco-Visible Mycoplasma Rapid Test Kit. https://www.mpbio.com/media/document/file/manual/dest/m/a/n/u/a/Manual_Mycoplasma_Rapid_Test_kit.pdf
49. Baczynska, A., Hvid, M., Lamy, P., Birkelund, S., Christiansen, G., Fedder, J. (2008). Prevalence of Mycoplasma genitalium, Mycoplasma hominis and Chlamydia trachomatis Among Danish Patients Requesting Abortion. Systems Biology in Reproductive Medicine, (54)3, 127–134. https://doi.org/10.1080/19396360802042838
50. Fisher Scientific. Drg International Inc Mycoplasma Hominis Igg Elisa. https://www.fishersci.com/shop/products/mycoplasma-hominis-igg-elisa/501947612#?keyword=ELISA%20mycoplasma%20hominis
51. Vitrotest ELISA kits & components. Urogenital Infections. https://vitrotest.ua/en/product-category/elisa/urogenital-infections
52. Hirschberg, L., Holme, T. (1991). ELISA for detection of Mycoplasma pneumoniae antigens using monoclonal antibodies. APMIS, (99), 475–481. https://doi.org/10.1111/j.1699-0463.1991.tb05178.x
53. Hama, H. O., Aboudharam, G., Barbieri, R. Lepidi, H., Drancourt, M. (2022). Immunohistochemical diagnosis of human infectious diseases: a review. Diagn Pathol, (2022)17:17. https://doi.org/10.1186/s13000-022-01197-5
54. Bahadır E. B., Sezgintürk M. K. (2016). Lateral flow assays: Principles, designs and labels. TrAC Trends in Analytical Chemistry, (82), 286–306 https://doi.org/10.1016/j.trac.2016.06.006
55. Miyashita, N., Kawai, Y., Tanaka, T., Akaike, H., Teranishi, H., Wakabayashi, T., Nakano, T., Ouchi, K., ..., Okimoto, N. (2015). Diagnostic sensitivity of a rapid antigen test for the detection of Mycoplasma pneumoniae: Comparison with real-time PCR. Journal of Infection and Chemotherapy. (21)6, 473–475. https://doi.org/10.1016/j.jiac.2015.02.007
56. Li W., Liu Y., Zhao Y., Tao, R., Li, Y., Shang, S. (2015). Rapid diagnosis of Mycoplasma pneumoniae in children with pneumonia by an immuno-chromatographic antigen assay. Sci Rep, 5, (15539)2015. https://doi.org/10.1038/srep15539
57. Yoon, SH., Min, IK., Ahn, JG. (2020). Immunochromatography for the diagnosis of Mycoplasma pneumoniae infection: A systematic review and meta-analysis. PLoS ONE, (15)3, e0230338. https://doi.org/10.1371/journal.pone.0230338
58. Singhal, N., Kumar, M., Kanaujia, P.K., Virdi, J.S. (2015). MALDI-TOF mass spectrometry: an emerging technology for microbial identification and diagnosis. Front Microbiol., (6-2015), 791. https://doi.org/10.3389/fmicb.2015.00791
59. Pailhoriès, H., Rabier, V., Eveillard, M., Mahaza, C., Joly-Guillou, M.-L. Chennebault, J.-M., Kempf, M., Lemarié, C. (2014). A case report of Mycoplasma hominis brain abscess identified by MALDI-TOF mass spectrometry. Int. J. Infect. Dis., (29), 166–168. https://doi.org/10.1016/j.ijid.2014.08.004
60. Reeve, M. A., Bachmann, D. (2019). MALDI-TOF MS protein fingerprinting of mixed samples. Biol. Methods Protoc., (4)1, bpz013. https://doi.org/10.1093/biomethods/bpz013
61. Chen, J., Huang, Y., Xiao, B., Deng, H., Gong, K., Li, K., Li, L., Hao, W. (2022). Development of a RPA-CRISPR-Cas12a Assay for Rapid, Simple, and Sensitive Detection of Mycoplasma hominis. Front. Microbiol., (13). https://doi.org/10.3389/fmicb.2022.842415
62. Ghouneimy, A., Mahas, A., Marsic, T., Aman, R., Mahfouz, M. (2023). CRISPR-Based Diagnostics: Challenges and Potential Solutions toward Point-of-Care Applications. ACS Synth. Biol., (12)1, 1‒16. https://doi.org/10.1021/acssynbio.2c00496.
63. Lonza. MycoAlert Mycoplasma Detection Kit. https://qb3.berkeley.edu/wp-content/uploads/2020/07/Mycoalert-protocol.pdf
64. Peerayeh, S. N., Samimi, R. (2008). Comparison of culture with the polymerase chain reaction for detection of gennital mycoplasma. European Journal of General Medicine, (5)2, 107-111. https://doi.org/10.29333/ejgm/82587
65. Keskin, F., Ciftci, S., Keceli, S.A., Koksal, M.O., Caliskan, E., Cakiroglu, Y., Agacfidan, A. (2018). Comparison of culture and real-time polymerase chain reaction methods for detection of Mycoplasma hominis in amniotic fluids samples. Nigerian Journal of Clinical Practice, (21)9, 1127–1131. https://doi.org/10.4103/njcp.njcp_230_17
66. Waites, K.B, Canupp K.C. (2001). Evaluation of BacT/ALERT System for Detection of Mycoplasma hominis in Simulated Blood Cultures. J Clin Microbiol, (39)12, 4328‒4331. https://doi.org/10.1128/jcm.39.12.4328-4331.2001
67. Allen-Daniels M. J., Serrano, M. G., Pflugner, L. P., Fettweis, J. M., Presto, M. A., Koparde, V. N., Brooks, J. P., Strauss, III, J. F., ..., Jefferson, K. K. (2015). Identification of a gene in Mycoplasma hominis associated with preterm birth and microbial burden in intraamniotic infection. Am. J. Obstet. Gynecol., (212)6, 779e1–779e13. https://doi.org/10.1016/j.ajog.2015.01.032
68. Waites, K.B., Xiao, L., Paralanov, V., Viscardi, R. M., Glass, J. I. (2012). Molecular Methods for the Detection of Mycoplasma and Ureaplasma Infections in Humans: A Paper from the 2011 William Beaumont Hospital Symposium on Molecular Pathology. J. Mol. Diagn., (14)5, 437–450. https://doi.org/10.1016/j.jmoldx.2012.06.001