Study on Drug Resistance to Treatment for Ureaplasma Urealyticum Infections in the Female Genital Tract.

BACKGROUND: Ureaplasma urealyticum (U. urealyticum) commonly occurs in female genitourinary infections, and its different biovars and serotypes have varying degrees of resistance to different antibiotics. This study aimed to ex-plore the characteristics of U. urealyticum infection and drug-resistant profiles in Chinese females. METHODS: We included 1,045 females with genital tract infections who visited Tangshan Workers' Hospital and Tangshan Maternal and Child Health Center from September 2017 to December 2018. The bacteria were selectively cultured, and drug sensitivity experiments were conducted. Eight pairs of oligonucleotide primers were designed, and polymerase chain reaction (PCR) was performed to amplify specific DNA fragments to perform bacterial strain typing. RESULTS: Among the 1,045 participants included, 566 (54.11%) participants were positive for mycoplasma infection. There were 432 (41.34%) participants with U. urealyticum infection, accounting for 76.33% of the positive participants. The infection rate of U. urealyticum was the highest in females who were 21 - 30 years old, followed by those who were 31 - 40 years old. Ureaplasma urealyticum showed the highest sensitivity to tetracyclines and the greatest resistance to quinolones. The biovar 1 of U. urealyticum with the highest detection rate of serotype 4, accounted for 66.88%. The biovar 2 of U. urealyticum mainly showed mixed subtypes 2 and 3. Biovar 2 showed higher resistance to sparfloxacin, clarithromycin, josamycin, and doxycycline than biovar 1. CONCLUSIONS: Women might be more susceptible to U. urealyticum, especially if they are of childbearing age. Urea-plasma urealyticum is mainly caused by a single serotype 6 infection. The resistance of U. urealyticum to quinolone (e.g., norfloxacin) is a great concern. Sparfloxacin, clarithromycin, ciprofloxacin, and doxycycline might be more suitable for people with biovar 1 infection. Biotyping may facilitate clinical drug use and help avoid the emergence of drug-resistant strains.

External quality assessment for molecular detection of Ureaplasma urealyticum in China.

BACKGROUND AND AIMS: A pilot external quality assessment (EQA) scheme for molecular detection of Ureaplasma urealyticum (UU) was conducted by the National Center for Clinical Laboratories (NCCL) to evaluate the testing capabilities of clinical laboratories and the actual performance of DNA-based nucleic acid amplification tests (NAAT) and RNA-based NAATs when applied in clinical settings. MATERIALS AND METHODS: The EQA panel contained twelve lyophilized samples, including positive samples containing inactivated cell culture supernatants of UU at different concentrations and sterile saline for negative samples. The positive samples were further divided into three groups of high, moderate and low concentrations. The panels were distributed to the participants and the datasets were analyzed according to the qualitative results. RESULTS: A total of 365 laboratories participated in the EQA scheme, and 360 results submitted by 338 laboratories were collected, of which 96.11 % (346/360) of the returned results and 95.86 % (324/338) of the laboratories were deemed competent. The positive percentage agreement (PPA) was ≥ 97.5 % for high and moderate concentration samples, but varied significantly for low concentration samples, decreasing from 86.94 % to 51.94 % as the sample concentration decreased. Additionally, for low concentration samples, RNA-based NAAT showed higher PPAs than DNA-based NAATs, but these results were specific to UU supernatants used in this study. CONCLUSION: Most of UU detection assays employed by the participants were generally consistent with their estimated limit of detection (LOD), and the majority of participants can reliably detect UU samples with high and moderate concentrations, while the poor analytical performance for low concentration samples requires further improvement and optimization.

Mycoplasma hominis increases the risk for Ureaplasma parvum infection in Human immunodeficiency virus infected pregnant women.

INTRODUCTION: Mycoplasma hominis and Ureaplasma parvum have been recently linked to sexually transmitted diseases and other conditions. There are a limited number of studies conducted on South African pregnant women that have assessed the prevalence and risk factors for genital mycoplasmas. METHODOLOGY: This study included 264 HIV infected pregnant women attending the King Edward VIII antenatal clinic in eThekwini, South Africa. DNA was extracted using the PureLink Microbiome kit and pathogens were detected using the TaqMan Real-time PCR assays. The statistical data analysis was conducted in a freely available Statistical Computing Environment, R software, version 3.6.3 using the RStudio platform. RESULTS: The prevalence of M. hominis and U. parvum, was 215/264 (81.4%), and 203/264 (76.9%), respectively. In the M. hominis positive group, a significantly (p = 0.004) higher proportion, 80.5% tested positive for U. parvum infection when compared to 61.2% among the M. hominis negative. Of the U. parvum positive women, a significantly (p = 0.004) higher proportion of women (85.2%) tested positive for M. hominis when compared to 68.9% among the U. parvum negative. In the unadjusted and adjusted analysis, being M. hominis positive increased the risk for U. parvum by approximately 3 times more (p = 0.014) and 4-fold (p = 0.008), respectively. CONCLUSIONS: This study showed a significant link between M. hominis and U. parvum infection. To date, there are a limited number of studies that have investigated M. hominisbeing a risk factor for U. parvum infection. Therefore, the data presented in the current study now fills in this gap in the literature.

Analyses of human papillomavirus, Chlamydia trachomatis, Ureaplasma urealyticum, Neisseria gonorrhoeae, and co-infections in a gynecology outpatient clinic in Haikou area, China.

BACKGROUND: The purpose of this study was to study the infection rates of Chlamydia trachomatis (CT), Ureaplasma urealyticum (UU), Neisseria gonorrhoeae (NG), and co-infections with human papillomavirus (HPV) in a hospital gynecology outpatient clinic in the Haikou region in 2021. METHODS: From January to December 2021, the Women and Children Medical Center of Hainan Province collected 2389 samples of cervical exfoliated cells and vaginal swab specimens from gynecologic outpatients. The samples were then analyzed descriptively for data, and the detection rate of each pathogen was tallied. All vaginal swabs were obtained for CT, UU, and NG DNA testing, and cervical exfoliated cells for HPV genotyping. Analyses were performed on the detection rate of each group. RESULTS: In 2389 samples, the frequencies of pathogen identification among the 2389 samples were as follows: UU (58.43%); HPV (17.29%); CT (7.99%); and NG (0.38%). HPV, CT, UU, and NG were detected in 33.33%, 22.55%, 77.45%, and 2.94% of individuals between 15 and 20 years of age, respectively. The detection rates of CT, UU, and NG were substantially greater in the HPV-positive group than the the HPV-negative group (P < 0.05). CONCLUSION: Among gynecologic outpatients at a hospital in the Haikou area, the probability of mixed infections with genital tract pathogens in HPV-positive patients was higher compared to HPV-negative patients. Reproductive tract infections are becoming more prevalent in younger people, hence adolescent sexual health education needs improvement.

Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum: hidden pathogens in peritoneal dialysis-associated peritonitis.

Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum commonly colonize the human urogenital tract, which may cause urogenital infections. However, infection by M. hominis, U. parvum, or U. urealyticum is rarely reported in patients with peritoneal dialysis (PD)-associated peritonitis. Herein, we reported four cases of PD-associated peritonitis caused by these pathogens, along with a review of the literature. The four cases were female patients with recurrent "culture-negative" PD-associated peritonitis and were related to menstruation. M. hominis, U. parvum, or U. urealyticum was detected in the PD fluid of the patients by metagenomic next-generation sequencing. All four patients were cured by intraperitoneal tigecycline combined with oral azithromycin or minocycline. M. hominis, U. parvum, and U. urealyticum should be paid more attention in female patients with recurrent culture-negative PD-associated peritonitis, especially when the peritonitis is related to menstruation, sexual intercourse, or urogenital tract operation. Moreover, metagenomic next-generation sequencing can provide a reasonable method to identify the pathogen for culture-negative PD-associated peritonitis.

A case of spondylodiscitis by Ureaplasma urealyticum in an immunocompetent adult with spine stabilization.

Ureaplasma urealyticum is part of the normal genital flora of many sexually experienced people, thereby it is mostly associated with genitourinary tract infections. Here, we present the first case reported in the literature of spondylodiscitis caused by U. urealyticum in a 62-year-old immunocompetent subject. U. urealyticum was detected through broad-range bacterial polymerase chain reaction in all samples obtained by T11 bone biopsy, while cultures were all negative. Due to the technical difficulties in removing the spinal osteosynthesis devices, no neurosurgical intervention was planned, therefore a suppressive therapy with moxifloxacin was administered. After 7 months, the patient underwent T10-11 partial vertebrectomy, insertion of an expandable cage at that level, the substitution of T11 screws, and prolongation of stabilization from T6 to ilium due to a fracture of T11 and T12; the remaining spinal osteosynthesis material was not removed. A computed tomography scan of the spine did not show features compatible with spondylodiscitis. Moxifloxacin was stopped after 15 months without any recurrence of U. urealyticum infection. Our case highlights the importance of considering U. urealyticum as a potential etiological germ in culture-negative spondylodiscitis.

DnaJ, a promising vaccine candidate against Ureaplasma urealyticum infection.

Ureaplasma urealyticum (U. urealyticum, Uu) is a common sexually transmitted pathogen that is responsible for diseases such as non-gonococcal urethritis, chorioamnionitis, and neonatal respiratory diseases. The rapid emergence of multidrug-resistant bacteria threatens the effective treatment of Uu infections. Considering this, vaccination could be an efficacious medical intervention to prevent Uu infection and disease. As a highly conserved molecular chaperone, DnaJ is expressed and upregulated by pathogens soon after infection. Here, we assessed the vaccine potential of recombinant Uu-DnaJ in a mouse model and dendritic cells. Results showed that intramuscular administration of DnaJ induced robust humoral- and T helper (Th) 1 cell-mediated immune responses and protected against genital tract infection, inflammation, and the pathologic sequelae after Uu infection. Importantly, the DnaJ protein also induced the maturation of mouse bone marrow-derived dendritic cells (BMDCs), ultimately promoting naïve T cell differentiation toward the Th1 phenotype. In addition, adoptive immunization of DnaJ-pulsed BMDCs elicited antigen-specific Immunoglobulin G2 (IgG2) antibodies as well as a Th1-biased cellular response in mice. These results support DnaJ as a promising vaccine candidate to control Uu infections. KEY POINTS: • A novel recombinant vaccine was constructed against U. urealyticum infection. • Antigen-specific humoral and cellular immune responses after DnaJ vaccination. • Dendritic cells are activated by Uu-DnaJ, which results in a Th1-biased immune response.

[Urogenital Ureaplasma urealyticum can cause invasive infection in immunosuppressed patients]. / Urogenital Ureaplasma gav invasiv infektion hos immunsupprimerad.

Ureaplasma urealyticum (Uu) is a bacterium without a cell wall, which makes it difficult to culture. Uu colonizes the lower genitourinary tract and is transmitted through sexual contact. The presence of Uu is higher in persons with immunosuppressive disease or treatment. Moreover, these persons are at increased risk of developing invasive Uu infections.  We present a case concerning a 47-year-old female with multiple sclerosis treated with Rituximab. She first presented with a urinary tract infection and bartholinitis. Despite treatment with antibiotics and surgical procedures, the infection disseminated and led to intra-abdominal abscesses and empyema. Repeated cultures were negative, which prolonged the time to diagnosis and accurate treatment. Uu was detected with 16S rRNA PCR assays during the course of the disease but was interpreted as non-pathogenic Finally, Uu was suspected as the causing agent, treatment with doxycycline was initiated, and the patient recovered after nine months of disease.

The effects of Chlamydia trachomatis, Mycoplasma hominis, and Ureaplasma urealyticum loads on semen quality: Detection and quantitative analysis.

BACKGROUND: The loads of Chlamydia trachomatis (CT), Mycoplasma hominis (MH), and Ureaplasma urealyticum (UU) may impact infertility, as well as cause risk of transmission. The quality and quantity of semen demonstrate male reproductive health. This study aimed to investigate the semen quality affected by CT, MH, and UU loads. MATERIALS AND METHODS: 130 semen samples, including infertile and fertile cases, were collected and analyzed. The whole genomic DNA was extracted, and the desired genes' plasmids were constructed. The CT, MH, and UU loads were quantified by real-time PCR. The data were analyzed using SPSS version 24. RESULTS: The average age of participants was 35.2 ± 6.8 years. CT, MH, and UU frequency were 9.2% vs. 3.1%, 15.4% vs. 3.1%, and 15.4 vs. 3.1% in infertile and fertile men, respectively. The mean loads of CT, MH, and UU in infertile men were 6.44 log10 copies/ml (range 5.31-7), 4.24 log10 copies/ml (range 3.37-4.7), and 6.94 log10 copies/ml (range 5.08-8.69) respectively, which was significantly higher than fertile men. The findings revealed a significant correlation between CT and UU loads and semen parameters, whereas the load of MH displayed significant effects just on sperm motility, morphology, and the number of leukocytes. CONCLUSION: The absence of clinical manifestations may not indicate the quality of semen. The pathogens' loads may significantly influence the quality and properties of male reproductive health.

Necrotizing funisitis associated with Ureaplasma urealyticum infection: A clinicopathologic analysis of 14 cases.

INTRODUCTION: Necrotizing funisitis is a distinct lesion of the umbilical cord associated with chorioamnionitis and bloodborne fetal infection. The lesion may be a response to microorganisms in Wharton's jelly. A common microorganism detected in chorioamnionitis is Ureaplasma urealyticum (U. urealyticum). This study hypothesizes that U. urealyticum DNA will be present in Wharton's jelly in necrotizing funisitis. METHODS: Necrotizing funisitis was identified retrospectively from a 2-year pathology database and confirmed in review. Paraffin fixed embedded tissue sections of the lesion were prepared for polymerase chain reaction (PCR) by using primers to identify U. urealyticum. Twenty matched controls without funisitis were similarly processed. Clinical data included serological tests of common bloodborne infections in the mothers and infants, and U. urealyticum PCR results in the urine of the neonates. RESULTS: Fourteen cases of necrotizing funisitis were identified in 7,416 examined placentas. Nine of these umbilical cords were positive by PCR for U. urealyticum (64.3%). Nineteen of twenty control cases were negative. Eight of ten neonates (80%) also had positive urine PCR tests for U. urealyticum. No infants or mothers had evidence of bloodborne fetal infection. DISCUSSION: U. urealyticum DNA was present in Wharton's jelly by PCR testing in the majority of the necrotizing funisitis lesions tested. This result supports a possible causative role for U. urealyticum in many cases of necrotizing funisitis.

Evaluation of Ureaplasma urealyticum, Chlamydia trachomatis, Mycoplasma genitalium and Neisseria gonorrhoeae in infertile women compared to pregnant women.

Infertility is one of the major health problems of patients suffering from bacterial infections. Given the high percentage of infertility, the aim of this study was to investigate the prevalence of Chlamydia trachomatis, Mycoplasma genitalium, Neisseria gonorrhoeae and Ureaplasma urealyticum in fertile and infertile women. In the prospective study, 65 infertile patients and 54 pregnant women referred to Mahdieh Hospital in Tehran were included. After transferring of vaginal swabs to the laboratory, DNA extraction and Polymerase Chain Reaction (PCR) were performed using specific primers. Of the 65 vaginal swab specimens, the prevalence of U. urealyticum, M. genitalium, C. trachomatis and N. gonorrhoeae were as 15 (23.1%), 11 (16.9%), 9 (13.8%) and 4 (6.2%), respectively; However, these rate in fertile group was as 6 (11.1%), 3 (5.5%), 5 (9.2%) and 1 (1.8%), respectively. Bacterial infections were higher in infertile group; therefore, these bacterial agents may be associated with female infertility. Timely control and treatment of infections caused by these organisms, together with other factors, can be important in prevention and treatment of the women's infertility and thereby community health.Impact StatementWhat is already known on this subject? Infertility is one of the most common reproductive health issues in Iran. Female reproductive system is a suitable environment for the growth of many pathogens, which may disrupt any stage of foetal formation, implantation or growth. Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium and Ureaplasma urealyticum are the most important microorganisms that have been considered in the infertility.What do the results of this study add? The prevalence of C. trachomatis, M. genitalium, N. gonorrhoeae, M. genitalium and U. urealyticum were higher in infertile women, but there was no statistically significant compared to pregnant women. These results suggest that timely control and treatment of infections caused by these organisms, along with other factors, can be used to prevent and treat women infertility and community health.What are the implications of these findings for clinical practice and/or further research? Based on the results, designing and implementing national control programs to prevent subsequent complications is thought to be necessary. Comprehensive analyses of the overall prevalence of these bacteria, particularly in developing countries (including Iran), may help to carry out such a strategy.

Propionibacterium acnes in urine and semen samples from men with urinary infection.

OBJECTIVE: Propionibacterium acnes has been implicated in the pathogenesis of prostate disease as acute and chronic prostatic inflammation, benign prostatic hyperplasia and prostate cancer although it should still be clarified if Propionibacterium acnes (P. acnes) is a commensal or accidental prostate pathogen. Aiming to evaluate the pathogenic potential for genitourinary tract of Propionibacterium acnes, we investigated the frequency of P. acnes genome in urine or semen samples from men with recurrent symptoms of urinary infection and negative testing for the most common urinary tract pathogens and sexually transmitted infections (STI) agents as Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum and Ureaplasma urealyticum. MATERIALS AND METHODS: The DNA extracted from urine and semen samples was analyzed for evaluating the P. acnes genome presence by real-time polymerase chain reaction (PCR). Infections were treated with vancomycin and cephalosporins antibiotics and then the search for the P.acnes genome by realtime PCR was repeated. RESULTS: The P. acnes qualitative real-time PCR revealed the genome in 73 out of 159 samples examined (108 urine and 51 semen). After antibiotic therapy, P. acnes was never detected. CONCLUSIONS: These results suggested that P. acnes genome determination should be performed in cases of chronic inflammation in the urinary tract to identify an unknown potential pathogen of genitourinary tract.

Development of multiplex real-time quantitative PCR for simultaneous detection of Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, and Mycoplasma genitalium in infertile women.

PURPOSE: Sexually Transmitted Diseases (STDs) can cause sterility and many other problems for women planning pregnancy. Currently, almost 340 million people worldwide suffer from Sexually Transmitted Infections (STIs). This study made attempts to quickly identify STDs' most critical infectious agents using dedicated primers and probes. METHODS: The present study was done on the cervical samples of 200 infertile women. After extracting the total DNA of Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, and Mycoplasma genitalium, quantitative methods were employed to determine the rate of target bacteria using multiplex real-time PCR. RESULTS: The multiplex qPCR showed the rates of 47%, 16%, 46%, and 16.5% for Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, and Mycoplasma genitalium in infertile women, respectively. In some patients, there were co-infections with two or three bacteria. The diagnostic approach used in our research could be employed as an alternative detection tool to identify the four most common STD-associated bacterial agents while detecting mixed infections. CONCLUSIONS: Infertile women with no biological problems could have their genital tract checked using this newly designed identification technique and get proper treatment for their infections as quickly as possible.

Diagnostic Value of Metagenomic Next Generation Sequencing for Ureaplasma urealyticum Infection: A Case Report.

Ureaplasma urealyticum has high nutritional requirements for culture, and it requires special tools for identification. Theoretically, metagenomic next generation sequencing (mNGS) can be used to detect many pathogens in clinical specimens, especially for complex infectious diseases with rare and atypical causes. Here, our patient developed severe pneumonia caused by U. urealyticum infection after allogeneic hematopoietic stem cell transplantation, and the etiology is unclear. After continuous negative culture, U. urealyticum was detected in the bronchoalveolar lavage fluid by mNGS, and azithromycin was used. Because of the difficulty in its diagnosis, diagnosis and treatment of extragenital U. urealyticum infection is challenging. In addition, many broad-spectrum antibiotics are ineffective against this pathogen because it lacks a cell wall. Therefore, early diagnosis and treatment are key to preventing further complications and deaths.

Diagnostic parameters of the AF Genital System® for detection of Mycoplasma hominis and Ureaplasma urealyticum.

OBJECTIVE: The prevalence of Mycoplasma hominis and Ureaplasma urealyticum (genital mycoplasma) amongst Indonesian women is poorly understood because of limited availability of diagnostic techniques. We sought to compare the diagnostic parameters of the AF Genital System® with those of culture methods and PCR as the gold standard for identification of M. hominis and U. urealyticum in vaginal swab specimens. METHODS: This was an observational diagnostic study. Eighty-eight specimens were collected from patients with abnormal vaginal discharge. Detection of M. hominis and U. urealyticum was performed using the AF Genital System®, culture methods, and PCR. RESULTS: Compared with PCR and culture methods, respectively, the AF Genital System® had sensitivities of 66.6% and 57% (M. hominis) and 55.5% and 77.8% (U. urealyticum). Compared with PCR and culture methods, respectively, the AF Genital System® had specificities of 82.9% and 86.5% (M. hominis) and 82.3% and 84.8% (U. urealyticum). CONCLUSION: The sensitivity of the AF Genital System® for detection of M. hominis and U. urealyticum from vaginal swab samples was lower than that of PCR, but specificity was reasonably good (82% to 83%).

Evaluation of the MYCOPLASMA IST3 urogenital mycoplasma assay in an international multicentre trial.

OBJECTIVES: To evaluate the accuracy, susceptibility and specificity of MYCOPLASMA IST3, the next generation of the most popular culture-based in vitro diagnostic device designed to detect, identify and test the susceptibility of urogenital mycoplasma infections. METHODS: MYCOPLASMA IST3 was evaluated against culture- and molecular-based gold standard methodologies to detect, identify, enumerate and determine antimicrobial resistance for Mycoplasma hominis and Ureaplasma species in 516 clinical samples collected across France, Serbia and the UK. Sample types included vulvovaginal/endocervical or urethral swabs (dry swab or eSwab®), semen and urine samples, which included blinded analysis following addition of a panel of 80 characterized control strains. RESULTS: Overall species identification was excellent for both Ureaplasma spp. (98.4% sensitivity, 99.7% specificity) and M. hominis (95.7% sensitivity, 100% specificity) relative to combined colony morphology on agar and quantitative PCR standards. Non-dilution-based bacterial load estimation by the assay was accurate between 83.7% (M. hominis) and 86.3% (Ureaplasma spp.) of the time (increased to 94.2% and 100%, respectively, if ±10-fold variance was allowed) relative to colonies counted on agar. Resistance accuracy for Ureaplasma spp. varied from gold standards for only 11/605 of individual tests (major error rate = 1.8%) and for 14/917 individual tests for M. hominis (major error rate = 1.5%). CONCLUSIONS: The redesigned MYCOPLASMA IST3 assay eliminated previous shortcomings by providing independent accurate resistance screening of M. hominis and Ureaplasma species, even in mixed infections, with CLSI-compliant thresholds. Specificity, sensitivity and enumeration estimates correlated closely with the confirmatory methods.

A New Multiplex Genetic Detection Assay Method for the Rapid Semi-Quantitative Detection of Six Common Curable Sexually Transmitted Pathogens From the Genital Tract.

Background: Sexually transmitted infections (STIs) are some of the most common communicable conditions and exert impact on the health and lives of many hundreds of millions of people across the world every year. Screening high-risk populations and conducting comprehensive detection tests would lead to a significant improvement in preventing the transmission of STIs and help us to provide rapid treatment to those affected. Here, we successfully established and validated a novel high-throughput multiplex gene detection system (HMGS) for the simultaneous and semiquantitative detection of six important curable sexually transmitted pathogens in a single reaction from secretions samples. Method: Fluorescently labeled primers were designed to target specific conserved and single-copy gene fragments of Ureaplasma urealyticum (U. urealyticum), Mycoplasma hominis (M. hominis), Chlamydia trachomatis (C. trachomatis), Neisseria gonorrhoeae (N. gonorrhoeae), Trichomonas vaginalis (T. vaginalis), and Treponema pallidum (T. pallidum). The specificity and sensitivity of the STI-HMGS was validated and optimized using plasmids and quantitative genomic DNA. Next, we validated the performances of the STI-HMGS for clinical application by testing samples of clinical secretions collected from patients who visited the gynecology and urology outpatient clinics of our reproductive medicine center. Results derived from the STI-HMGS were then compared with three approved commercialized kits that used to detect U. urealyticum, C. trachomatis and N. gonorrhoeae, respectively, followed by further validation with Sanger sequencing for all pathogens. Finally, a comprehensive analysis of epidemiology was performed among different subgroups to investigate the association between infection rates and clinically-relevant information. Results: The sensitivity of STI-HMGS for six target genes was 10 copies/µL. Data derived from the detection of 381 clinical secretions demonstrated that the STI-HMGS exhibited high concordance rate compared with approved commercialized kits and almost 100% sensitivity and specificity for the detection of six sexually transmitted pathogens when validated by Sanger sequencing. Semi-quantitative analysis found that STIs caused by N. gonorrhoeae had a significantly higher (P<0.05) pathogen load than the other pathogens. Infections caused by C. trachomatis were significantly more common in younger individuals (P<0.05). We also found that U. urealyticum infections were more likely to happen in females; while the males were more affected by N. gonorrhoeae (P<0.05). Conclusions: STI-HMGS proved to be an efficient method for the semi-quantitative detection of six important curable sexually transmitted pathogens and therefore represents an alternative method for the clinical detection and monitoring of STIs.

Droplet digital PCR (ddPCR) for the detection and quantification of Ureaplasma spp.

BACKGROUND: Ureaplasma spp. are associated with various infectious diseases in females, but there is still limited evidence regarding whether they are related to nonspecific cervicitis. The aim of this study was to develop and evaluate a digital droplet PCR (ddPCR) assay for the detection and quantification of Ureaplasma spp. in cervical swabs. METHODS: A total of 267 non-specific cervicitis (NSC) patients and 195 asymptomatic females were included in this study. We produced standard curves for Ureaplasma spp. to evaluate the analytical performance of the ddPCR assay. Then, we detected and quantified the bacterial load of Ureaplasma spp. in cervical swabs. RESULTS: The prevalences of U. parvum were 37.8% (101/267) and 29.7% (58/195),  U. urealyticum were 9.0% (24/267) and 8.7% (17/195) in the NSC group and control group, respectively. In addition, the median copy number of U. parvum was 2.5 × 104 copies/ml (n = 101) in the NSC group and 9.2 × 103 copies/ml (n = 58) in the control group. The U. parvum load in the NSC group was significantly higher than that in the asymptomatic individuals (P < 0.001). whereas the median load of U. urealyticum was 8.4 × 103 copies/ml (n = 24) and 1.4 × 103 (n = 17) copies/ml in the two groups, respectively, , the difference was not statistically significant (P = 0.450). CONCLUSIONS: Our study is the first to develop a droplet digital PCR (ddPCR) method for the detection and quantification of Ureaplasma spp. in clinical samples, and the method has excellent analytical performance and a wide range of clinical application prospects.

[Comparison of two methods for detection of Chlamydia trachomatis and Ureaplasma urealyticum in male reproductive tract].

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.