This study primarily aimed to investigate the prevalence of human papillomavirus (HPV) and other common pathogens of sexually transmitted infections (STIs) in spermatozoa of infertile men and their effects on semen parameters. These pathogens included Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae, Pseudomonas aeruginosa , and Staphylococcus aureus . A total of 1951 men of infertile couples were recruited between 23 March 2023, and 17 May 2023, at the Department of Reproductive Medicine of The First People's Hospital of Yunnan Province (Kunming, China). Multiplex polymerase chain reaction and capillary electrophoresis were used for HPV genotyping. Polymerase chain reaction and electrophoresis were also used to detect the presence of other STIs. The overall prevalence of HPV infection was 12.4%. The top five prevalent HPV subtypes were types 56, 52, 43, 16, and 53 among those tested positive for HPV. Other common infections with high prevalence rates were Ureaplasma urealyticum (28.3%), Ureaplasma parvum (20.4%), and Enterococcus faecalis (9.5%). The prevalence rates of HPV coinfection with Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae , and Staphylococcus aureus were 24.8%, 25.4%, 10.6%, 6.4%, 2.4%, 7.9%, 5.9%, 0.9%, and 1.3%, respectively. The semen volume and total sperm count were greatly decreased by HPV infection alone. Coinfection with HPV and Ureaplasma urealyticum significantly reduced sperm motility and viability. Our study shows that coinfection with STIs is highly prevalent in the semen of infertile men and that coinfection with pathogens can seriously affect semen parameters, emphasizing the necessity of semen screening for STIs.
Humans ; Male ; Infertility, Male/epidemiology* ; Coinfection/microbiology* ; Papillomavirus Infections/virology* ; Adult ; Sexually Transmitted Diseases/complications* ; China/epidemiology* ; Staphylococcus aureus/isolation & purification* ; Chlamydia trachomatis/isolation & purification* ; Prevalence ; Mycoplasma genitalium/isolation & purification* ; Ureaplasma urealyticum/isolation & purification* ; Neisseria gonorrhoeae/isolation & purification* ; Enterococcus faecalis/isolation & purification* ; Streptococcus agalactiae/isolation & purification* ; Herpesvirus 2, Human/genetics* ; Pseudomonas aeruginosa/isolation & purification* ; Semen/virology* ; Sperm Motility ; Spermatozoa/microbiology* ; Human Papillomavirus Viruses

Objective To make the bioinformatics analysis of Ureaplasma parvum UP3-RS02445 and prepare monoclonal antibody (mAb) against UP3-RS02445. Methods The biological characteristics of UP3-RS02445 protein were predicted by bioinformatics software. The UP3-RS02445 prokaryotic expression plasmid was constructed and the corresponding protein expression was induced by isopropyl-β-D-thiogalactoside (IPTG). Thus the expressed protein was used as immunogen to immunize female BALB/c mice. Hybridoma cell technology was used to prepare the monoclonal antibody against UP3-RS02445. The specificity and titer of monoclonal antibody were detected by Western blot and ELISA respectively. The subclass of heavy chain and subtype of light chain were identified by monoclonal antibody subtype identification test strip. Results Bioinformatics analysis showed that UP3-RS02445 protein was composed of 201 amino acids, without transmembrane domain and signal peptide, and belongs to non-secretory proteins. The recombinant prokaryotic plasmid of UP3-RS02445 was successfully constructed and the recombinant protein could be induced in large amount. After cell fusion, two hybridoma cells (A1H5 and A4E2) secreting UP3-RS02445 mAb were screened by ELISA and Western blot. The results of ELISA showed that the titers of monoclonal antibodies were 1:2560. Western blot and Immunofluorescence technique both indicated that the antibodies could bind specifically to the UP3-RS02445 protein. The heavy chain and light chain of the two mAbs were IgG1 and kappa subtypes respectively. Conclusion We prepared the UP3-RS02445 monoclonal antibodies with well specificity and high titer which might lay foundations for the subsequent development of UP diagnostic reagents and the functional study of protein.
Antibodies, Monoclonal/immunology* ; Animals ; Mice, Inbred BALB C ; Female ; Computational Biology/methods* ; Mice ; Ureaplasma urealyticum/genetics* ; Bacterial Proteins/genetics* ; Antibody Specificity ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology*

OBJECTIVE: To investigate the value of clinical application of simultaneous amplification and testing of RNA (SAT-RNA) for detecting Chlamydia trachomatis (CT) and Ureaplasma urealyticum (UU) by comparing with the polymerase chain reaction testing of DNA (PCR-DNA) method. METHODS: Specimens from both urethra swab and the first avoid urine which should be at least one hour after the previous urination were collected from 163 men who were scheduled for in vitro fertilization and embryo transfer (IVF-ET) treatment due to female factors at Center for Reproductive Medicine, Shengjing Hospital of China Medical University during the period of April 2016 to April 2017. Among the 163 men, 109 simultaneously provided semen that was collected after 3-7 days of sexual abstinence for the testing. Urine and semen specimens were detected for CT and UU with SAT-RNA, while urethra swab specimens were detected for CT and UU with standard PCR-DNA. Detection results of the SAT-RNA were compared with those of the PCR-DNA method. RESULTS: The positive rate of UU in the urethra swab detected with PCR-DNA and that of UU in the urine with SAT-RNA were 47.24% and 47.85%, respectively, and the coincidence rate was 93.25%. In addition, the positive and negative coincidence rates were 93.51% and 93.02%, respectively, and the concordance between the two methods was very good (Kappa=0.865). On the other hand, the positive rate of CT in the swab specimen tested with PCR-DNA was 3.07% and that of CT in urine with SAT-RNA was 4.29%, and the coincidence rate was 97.55%. Moreover, the positive and negative coincidence rates were 80.00% and 98.10%, respectively, and the concordance between the two methods was good (Kappa=0.654). Regarding SAT-RNA detection of UU in the urine and semen specimen of the 109 patients, the positive rates of UU in the urine and semen specimens were 50.46% and 44.95%, respectively; and the coincidence rate between the two specimens was 88.99%. In addition, the positive coincidence rate and the negative coincidence rate was 93.88% and 85.00%, respectively, and the concordance between the two specimens was good (Kappa=0.780). Similarly, SAT-RNA detection of CT in the urine and semen specimens showed the positive rate was 5.50% and 3.67%, respectively; and the two specimens showed 98.17% coincidence rate. The positive and negative coincidence rates were 100.00% and 98.10%, respectively, and the concordance was also good (Kappa=0.791). CONCLUSION SAT-RNA detection of CT and UU in the urine specimen showed good concordance with the PCR-DNA detection of CT and UU in the urethra swab specimen. In addition, the concordance was also good between the urine and semen specimens detected with SAT-RNA. These results indicate that, as a less invasive and equally accurate procedure, SAT-RNA may be more suitable for clinical application.
Chlamydia Infections/epidemiology* ; Chlamydia trachomatis/genetics* ; Female ; Humans ; Infertility, Male ; Male ; Neisseria gonorrhoeae/genetics* ; Polymerase Chain Reaction ; Ureaplasma urealyticum/genetics*

Objective: To investigate the value of real-time RNA simultaneous amplification and testing (SAT) in the detection of Ureaplasma urealyticum (UU) in the semen of infertile males and its clinical significance. METHODS: We collected semen samples from 542 infertility patients and 120 normal fertile men as controls in the Andrology Clinic of Nanjing General Hospital from March to September 2015. We detected UU infection in the samples using the culture method and SAT technology, respectively. RESULTS: All the UU positive cases (except 4 false positive cases) detected by the culture method were also shown to be positive in SAT. The UU detection rate of SAT was significantly higher than that of the culture method both in the infertility patients (54.1 vs 19.7%, P<0.05) and in the normal controls (42.5 vs 12.5%, P<0.05). CONCLUSIONS SAT is a rapid and accurate method for detecting UU infection in semen samples, with a higher sensitivity and accuracy than the culture method, and it can also be used to evaluate the therapeutic effects. However, the culture method has its own advantages, such as low requirement of technical equipment, easy operation, and possibility of drug sensitivity test at the same time. Therefore, SAT and the culture method can be used alternatively according to the clinical need.
Andrology ; Humans ; Infertility, Male ; microbiology ; Male ; Nucleic Acid Amplification Techniques ; RNA, Bacterial ; analysis ; Semen ; chemistry ; microbiology ; Semen Analysis ; Ureaplasma Infections ; diagnosis ; Ureaplasma urealyticum ; genetics ; isolation & purification

Chlamydia trachomatis ; genetics ; Chlamydiaceae Infections ; diagnosis ; genetics ; Genes, Bacterial ; genetics ; Genes, Viral ; genetics ; Gonorrhea ; diagnosis ; genetics ; Herpes Simplex ; diagnosis ; genetics ; Herpesvirus 2, Human ; genetics ; Humans ; Multiplex Polymerase Chain Reaction ; Neisseria gonorrhoeae ; genetics ; Ureaplasma Infections ; diagnosis ; genetics ; Ureaplasma urealyticum ; genetics

OBJECTIVETo study the relationship between microbial gene 16SrRNA and intrauterine infection.

METHODSThirty cases of single preterm birth were enrolled, including 16 cases due to premature rupture of membranes (PROM) (rupture time>18 hrs), 6 cases due to spontaneous preterm birth and 8 cases due to iatrogenic preterm birth. Ten cases of single term birth were used as the control group. Fetal membrane and placenta samples were obtained. Amniotic fluid, blood from cord or newborn babies as well as gastric fluid and tracheal secretions from infants with mechanical ventilation were also obtained. The histological features of placenta and fetal membranes were observed. Polymerase chain reaction (PCR) was used to detect the presence of microbial 16SrRNA and ureaplasma urealyticum (UU) in placenta, fetal membranes and other samples.

RESULTSTwenty-one (70%) cases were diagnosed as chorioamnionitis, characterized by neutrophil infiltration in fetal membrane and placenta tissues, especially in fetal membranes. Chorioamnionitis was most frequent in babies whose gestational age less than 32 weeks or birth weight lower than 1 500 g. Positive 16SrRNA gene was found in 12 cases, and positive UU gene in 10 cases in the preterm birth group. Neither 16SrRNA nor UU gene was detected in the control group. The PROM preterm babies developed more frequent infection than the babies premature born due to other causes, but there were no statistically significant differences in the incidence of infection.

CONCLUSIONSChorioamnionitis may be the major cause of PROM and premature birth. The detection of microbial genes is valuable in identification of intrauterine infection.

Chorioamnionitis ; diagnosis ; Female ; Fetal Membranes, Premature Rupture ; etiology ; Humans ; Infant, Newborn ; Infant, Premature ; Placenta ; microbiology ; pathology ; Pregnancy ; RNA, Ribosomal, 16S ; genetics ; Ureaplasma urealyticum ; genetics ; isolation & purification

The aim of this study was to evaluate the occurrence of genital mycoplasmas, especially Ureaplasma parvum and Ureaplasma urealyticum, in women with atypical squamous cells of undetermined significance (ASCUS), low grade squamous intraepithelial lesions (LSIL) and high grade squamous intraepithelial lesions (HSIL), compared to women with normal cytology living in Katowice, Poland. Two sterile swabs were used to obtain material from the posterior vaginal fornix of 143 women with squamous intraepithelial lesions and 39 healthy women: first for general bacteriology, second for detection of urogenital mycoplasmas using Mycoplasma IST2 kit. From each positive Mycoplasma IST2 culture DNA was isolated and PCR was performed for identification of U. parvum and U. urealyticum. Mycoplasma IST was positive in 34.1% cases. Urogenital mycoplasmas were demonstrated in women with HSIL significantly more often compared to women with LSIL, ASCUS, and with normal cytology. DNA of U. parvum was demonstrated in majority of Mycoplasma IST2-positive cases, U. urealyticum DNA-only in 9 (4.9%). Predominance of 3/14 serovars of U. parvum was demonstrated. U. urealyticum biovar 2 was present more often in women with squamous intraepithelial lesions.
Adult ; Animals ; Female ; Humans ; Male ; Poland/epidemiology ; Pregnancy ; Ureaplasma/*genetics ; Ureaplasma Infections/epidemiology/*microbiology ; Ureaplasma urealyticum/*genetics ; Uterine Cervical Dysplasia/*microbiology/pathology

OBJECTIVETo investigate the correlation between Ureaplasma urealyticum infection and infertility in Chinese males.

METHODSAccording to the results of the heterogeneity test, a comprehensive quantitative analysis was made of 49 papers on Ureaplasma urealyticum infection and Chinese male infertility by RevMan 4.2.2. The impacts of different sample volumes on the research findings were compared, and the sensitivities of culture and PCR detections analyzed respectively.

RESULTSUreaplasma urealyticum had a significant negative impact on Chinese male fertility. Based on different samples of literature, two rounds of screening and analysis were carried out and two different conclusions derived. The first was OR = 4.43 (95% CI: 3.77-5.22), with the OR values of culture and PCR detections as 4.25 (95% CI: 3.59-5.03) and 5.35 (95% CI: 3.37-8.47), and the second was OR = 4.28 (95% CI: 3.52-5.20), with the OR values of culture and PCR detections as 4.24 (95% CI: 3.41-5.28) and 4.42 (95% CI: 2.73-7.17).

CONCLUSIONThere is a significant correlation between Ureaplasma urealyticum and Chinese male infertility. The conclusion of study is significantly influenced by the sample volume, which should be reasonably designed. The sensitivity of PCR detection is higher than that of culture detection.

China ; epidemiology ; Humans ; Infertility, Male ; epidemiology ; Male ; Polymerase Chain Reaction ; Ureaplasma Infections ; epidemiology ; microbiology ; Ureaplasma urealyticum ; genetics ; isolation & purification

OBJECTIVETo study and compare the pathogenicity and pathogenic condition of Ureaplasma urealyticum (Uu) and Mycoplasma homonis (Mh) between healthy women and women with gential tract inflammation.

METHODSTo collect the gential tract secretion in the two groups and detect the infectious ratio and color change unit ( CCU) concentration. Together with data gathered from questionnaires, we studied the mycoplasma infectious status between the two kinds of people.

RESULTSThe positive rate was 76. 1 % in women with gential inflammation, higher than in healthy women whose positive rate was 42.2% (chi(2) = 45.1862, P< 0.0001). Mixed infection of Uu and Mh was popular in infected women. Healthy women were easier to be infected by Uu or Mh( Uu, Uu + Mh: X(2) = 39.5956, P< 0.0001; Mh,Uu + Mh: X(2)= 13.2935, P= 0.0003). The result of CCU concentration showed the infected concentration in women with gential tract inflammation was higher than healthy women(Uu: Z = 7. 1058, P< 0.0001; Mh: Z= 8.7201, P< 0.0001). Uu and Mh were commonl sensitively in every age.

CONCLUSIONBoth Uu and Mh were conditioned pathogens. The two kinds of mycoplasma had cooperated pathogenic effects which was easily leading to clinical symptom in the high infectious concentration.

Adult ; Aged ; China ; epidemiology ; Female ; Genital Diseases, Female ; epidemiology ; immunology ; microbiology ; Humans ; Middle Aged ; Mycoplasma ; genetics ; isolation & purification ; Mycoplasma Infections ; epidemiology ; microbiology ; Ureaplasma Infections ; epidemiology ; microbiology ; Ureaplasma urealyticum ; genetics ; isolation & purification ; Women's Health ; Young Adult

OBJECTIVETo establish the approach to detecting two biovars of Ureaplasma urealyticum (Uu) in human semen and to investigate the relationship between the two biovars of Uu infection and the quality of human semen.

METHODSBased on the 16S-23S rRNA intergenic spacer region, three pairs of primers were designed, the species specific primer and two biovars primers (Parvo primer and T960 primer). The two biovars of Uu were detected in the semen from 949 men by semen culture and PCR assay. Meanwhile, semen routine analyses were performed.

RESULTSIn the 949 subjects, 199 were Uu positive both in Uu liquid culture and PCR assay (199/949, 21.1%), of which 136 (136/199, 68.3%) were Parvo biovar, 54 (54/199, 27.1%) T960 biovar, and 9 (9/199, 4.5%) both Parvo and T960 biovars. Compared with the Parvo and the negative groups, human sperm viability was significantly decreased (P < 0.05 ) in the Uu T960 infection group. The difference of sperm motility and density had no statistic significance.

CONCLUSIONA significant correlation has been found between Uu T960 biovar infection and human sperm viability

Adult ; Humans ; Male ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; genetics ; RNA, Ribosomal, 23S ; genetics ; Semen ; microbiology ; Sperm Motility ; Ureaplasma urealyticum ; classification ; genetics