Diagnostic capabilities of ejaculate sediment examination with the help of nucleic acid amplification by polymerase chain reaction and microbiologic examination in searching for causes of male reproductive disfunction

Sapozhkova Zh.Yu.; Milovanova G.A.; Pochernikov D.G.; Postovoitenko N.T.

doi:https://doi.org/10.17116/medtech20224402162

Sapozhkova Zh.Yu.

International Cytology School and Medical School of Innovations;
Podolsk Diagnostic Center LLC

Milovanova G.A.

Podolsk Diagnostic Center LLC;
Russian Medical Academy of Postgraduate Professional Education

Pochernikov D.G.

Ivanovo State Medical Academy

Postovoitenko N.T.

Mother and Child Vladimir LLC

Diagnostic capabilities of ejaculate sediment examination with the help of nucleic acid amplification by polymerase chain reaction and microbiologic examination in searching for causes of male reproductive disfunction

Authors:

Sapozhkova Zh.Yu., Milovanova G.A., Pochernikov D.G., Postovoitenko N.T.

More about the authors

Journal: Medical Technologies. Assessment and Choice. 2022;44(2): 62‑71

DOI: 10.17116/medtech20224402162

Read: 1744 times

Download PDF (RU)

Contact the author

Table of contents

To cite this article:

Sapozhkova ZhYu, Milovanova GA, Pochernikov DG, Postovoitenko NT. Diagnostic capabilities of ejaculate sediment examination with the help of nucleic acid amplification by polymerase chain reaction and microbiologic examination in searching for causes of male reproductive disfunction. Medical Technologies. Assessment and Choice. 2022;44(2):62‑71. (In Russ.)
https://doi.org/10.17116/medtech20224402162

Подписка 2026 Медицинские технологии. Оценка и выбор (Medical Technologies. Assessment and Choice)

Использование инфографики авторами научных работ

Close metadata

OBJECTIVE

To analyze the diagnostic capabilities of nucleic acid amplification (NAA) by real-time polymerase chain reaction (PCR) and microbiological examination of ejaculate sediment in determining the causes of male reproductive dysfunction.

MATERIAL AND METHODS

We examined 529 samples of ejaculate sediment from men aged 18—59 years with infertility and male accessory gland infection (Male Accessory Gland Infection/MAGI) between 2017 and 2021. Microorganisms for microbiological and molecular analysis were identified by matrix-assisted laser desorption/ionization, time-of-flight mass spectrometry (MALDI-TOF MS) and real-time PCR (Androflor test, NAAT/PCR-RV/Androflor).

RESULTS

As a rule, significant bacteriospermia in ejaculate sediment was detected in Androflor molecular test compared to microbiological examination (402 (76%) and 237 (44.8%), respectively, p<0.01). Analysis of biotope by two methods indicates complete coincidence of microbial status only in 40 (7.6%) out of 529 cases. Otherwise, microbiological examination found no microbiota (83 (15.7%) sample) or spectrum of detected microorganisms differed. There were differences in the structure of isolated groups of microorganisms in cluster of predominant obligate anaerobic conditionally pathogenic microorganisms (p<0.01), gram-positive facultative anaerobic microorganisms Pseudomonas aeruginosa/Ralstonia spp./Burkholderia spp. (p<0.01), Haemophilus spp. (p<0.01) and monoculture Enterobacteriaceae spp./Enterococcus spp. (p<0.01). In the group with normal flora, NAA/RT-PCR/Androflor test revealed common associations of microorganisms (103 (81.1%)), microbiological examination revealed monoculture in most cases (216 (74%)) (p<0.01).

CONCLUSION

The Androflor molecular test is recommended for effective laboratory examination and timely treatment of men with reproductive dysfunction. Microbiological examination is also important since this method is valuable to analyze sensitivity of microorganisms to antibiotics and detect various enterobacteria requiring different therapeutic regimens.

Keywords:

ejaculate sediment analysis

RT-PCR

Androflor

microbiological analysis

male accessory gland infection

male infertility

Authors:

Sapozhkova Zh.Yu.

International Cytology School and Medical School of Innovations;
Podolsk Diagnostic Center LLC

Milovanova G.A.

Podolsk Diagnostic Center LLC;
Russian Medical Academy of Postgraduate Professional Education

Pochernikov D.G.

Ivanovo State Medical Academy

Postovoitenko N.T.

Mother and Child Vladimir LLC

Received:

21.02.2022

Accepted:

25.03.2022

Close metadata

References:

WHO Health Topics. Infertility. Accessed January 28, 2022. https://www.who.int/health-topics/infertility#tab=tab_1
Agarwal A, Mulgund A, Hamada A, Chyatte MR. A unique view on male infertility around the globe. Reproductive Biology and Endocrinology: RB&E. 2015;13:37. https://doi.org/10.1186/s12958-015-0032-1
Levine H, Jørgensen N, Martino-Andrade A, Mendiola J, Weksler-Derri D, Mindlis I, et al. Temporal trends in sperm count: a systematic review and meta-regression analysis. Human Reproduction Update. 2017;23(6):646-659. https://doi.org/10.1093/humupd/dmx022
Moretti E, Figura N, Campagna MS, Iacoponi F, Gonnelli S, Collodel G. Infectious Burden and Semen Parameters. Urology. 2017;100:90-96. https://doi.org/10.1016/j.urology.2016.10.032
Ahmadi MH, Mirsalehian A, Sadighi Gilani MA, Bahador A, Talebi M. Asymptomatic Infection with Mycoplasma hominis Negatively Affects Semen Parameters and Leads to Male Infertility as Confirmed by Improved Semen Parameters After Antibiotic Treatment. Urology. 2017;100:97-102. https://doi.org/10.1016/j.urology.2016.11.018
Agarwal A, Parekh N, Panner Selvam MK, Henkel R, Shah R, Homa ST, et al. Male Oxidative Stress Infertility (MOSI): Proposed Terminology and Clinical Practice Guidelines for Management of Idiopathic Male Infertility. The World Journal of Men’s Health. 2019;37(3):296-312. https://doi.org/10.5534/wjmh.190055
Voroshilina ES, Zornikov DL, Panacheva EA. Comparative study of the microbiota of ejaculate by quantitative PCR and culture method. Vestnik Rossijskogo gosudarstvennogo medicinskogo universiteta. 2019;1:44-49. (In Russ.). https://doi.org/10.24075/vrgmu.2019.009
Pochernikov DG, Postovoitenko NT, Strelnikov AI. Comparative analysis of cultural and molecular genetic methods in the study of the microbiota of ejaculate in male infertility. Andrologiya i genital’naya hirurgiya. 2019;20(2):40-47. (In Russ.). https://doi.org/10.17650/2070-9781-2019-20-2-40-47
Voroshilina ES, Zornikov DL, Ivanov AV, Pochernikov DG, Panacheva EA. Microbiota of ejaculate: cluster analysis of the results obtained by PCR-RV. Vestnik Rossijskogo gosudarstvennogo medicinskogo universiteta. 2020;5:66-73. (In Russ.).
WHO Laboratory Manual for the Examination and Processing of Human Semen. Sixth edition. 2021. Accessed January 28, 2022. https://www.who.int/publications/i/item/9789240030787
Instrukciya po primeneniyu nabora reagentov dlya issledovaniya mikroflory urogenital’nogo trakta muzhchin metodom PCR v rezhime real’nogo vremeni. Fasovka standartnaya (S). Varianty komplektacii: Androflor i Androflor Skrin (OOO NPO «DNK-Tekhnologiya», Rossiya). Accessed January 28, 2022. (In Russ.). https://www.dna-technology.ru/sites/default/files/393-4_s_androflorskrin_ivd_b_2021-06-21_0.pdf EAU
Guidelines on Urological Infections. 2022. Accessed January 28, 2022. https://d56bochluxqnz.cloudfront.net/documents/full-guideline/EAU-Guidelines-on-Urological-Infections-2022.pdf
EAU Guidelines on Male Infertility. 2019. Accessed January 28, 2022. https://d56bochluxqnz.cloudfront.net/media/EAU-Guidelines-on-Male-Infertility-2019.pdf
Muzhskoe besplodie. Kliniheskie rekomendacii. Odobreny Nauchno-prakticheskim Sovetom Minzdrava Rossii. 2021. (In Russ.).
Dermatovenerologiya 2015: Bolezni kozhi. Infekcii, peredavaemye polovym putem. Federal’nye klinicheskie rekomendacii. M.: Delovoj ekspress; 2016. Accessed January 28, 2022. (In Russ.). https://cnikvi.ru/docs/2335_maket_30.pdf
Cregger M, Lenz KM, Leary E, Leach RE, Fazleabas AT, White BA, et al. Reproductive Microbiomes: Using the Microbiome as a Novel Diagnostic Tool for Endometriosis. Reproductive Immunology: Open Access. 2017;2(3):36. https://doi.org/10.21767/2476-1974.100036
Mändar R, Punab M, Borovkova N, Lapp E, Kiiker R, Korrovits P, et al. Complementary seminovaginal microbiome in couples. Research in Microbiology. 2015;166(5):440-447. https://doi.org/10.1016/j.resmic.2015.03.009
Tao X, Franasiak JM, Zhan Y, Scott RT, Rajchel J, Bedard J, et al. Characterizing the endometrial microbiome by analyzing the ultra-low bacteria from embryo transfer catheter tips in IVF cycles: Next generation sequencing (NGS) analysis of the 16S ribosomal gene. Human Microbiome Journal. 2017;3:15-21. https://doi.org/10.1016/J.HUMIC.2017.01.004
Freitas AC, Chaban B, Bocking A, Rocco M, Yang S, Hill JE, et al.; VOGUE Research Group. The vaginal microbiome of pregnant women is less rich and diverse, with lower prevalence of Mollicutes, compared to non-pregnant women. Scientific Reports. 2017;7(1):9212. https://doi.org/10.1038/s41598-017-07790-9
Budd WT, Harwich MD, Meyers G, Dilts JR, Woody JR, Reynolds T. Metagenomics Analysis Using Next Generation Sequencing of Vaginal Samples from Community Practices in the US. MOJ Cell Science and Report. 2015;2(2):16-25. https://doi.org/10.15406/mojcsr.2015.02.00020
García-Velasco JA, Menabrito M, Catalán IB. What fertility specialists should know about the vaginal microbiome: a review. Reproductive Biomedicine Online. 2017;35(1):103-112. https://doi.org/10.1016/j.rbmo.2017.04.005
Sapozhkova ZhYu. Sposob mikroskopicheskoj diagnostiki kachestva spermy posle sedimentacii eyakulyata. Patent RF na izobretenie №2686685/30.04.2019. Byul. №13. Accessed January 28, 2022. (In Russ.). https://test2.fips.ru/ofpstorage/Doc/IZPM/RUNWC1/000/000/002/686/685/%D0%98%D0%97-02686685-00001/DOCCLAIM.PDF
Sapozhkova Z, Kasoyan K, Kovalchuk E, Shabalova I. Sperm Sediment Cytology: A New Technique for Diagnosing Occult Urologic Infections. Acta Cytologica. 2017;61(3):247-251. https://doi.org/10.1159/000469653
Sapozhkova ZhYu, Milovanova GA, Patsap OI. Laboratory diagnostics of male infertility. Biomarkers and methods. Part II. Laboratornaya i klinicheskaya medicina. Farmaciya. 2021;1(2):65-79. (In Russ). https://doi.org/10.14489/lcmp.2021.02.pp.065-079
Sapozhkova ZhYu, Shabalova IP, Kasoyan KT. Issledovanie osadka eyakulyata v diagnostike infekcii, peredavaemyh polovym putem: uchebnoe posobie. M.—Tver’: Triada; 2017. (In Russ.).
Sapozhkova ZhYu, Milovanova GA, Patsap OI. Laboratory diagnostics of male infertility. Biomarkers. Part I. Laboratornaya i klinicheskaya medicina. Farmaciya. 2021;1(1):57-68. (In Russ). https://doi.org/10.14489/lcmp.2021.01.pp.057-068
Jarvi K, Lacroix JM, Jain A, Dumitru I, Heritz D, Mittelman MW. Polymerase chain reaction-based detection of bacteria in semen. Fertility and Sterility. 1996;66(3):463-467.