RESUMO
Introduction: This study aims to determine the etiological, sociodemographic, and clinical characteristics of STIs, and the level of resistance in M. genitalium in Shenzhen, a representative first-tier city of southern China. Methods: A multicenter cross-sectional study was conducted and 7886 sexually active participants attending STI-related departments were involved from 22 hospitals. Nine STI-related organisms including N. gonorrhoeae, C. trachomatis, T. vaginalis, M. genitalium, HSV-1, HSV-2, M. hominis, U. parvum, and U. urealyticum were screened. Results: Being single or divorced was associated with increased detection of N. gonorrhoeae, C. trachomatis, M. genitalium, HSV-1, HSV-2 and M. hominis. Lower education level was associated with increased detection of C. trachomatis, HSV-2 and M. hominis. No insurance coverage was an independent risk factor for T. vaginalis, M. hominis and U. parvum positivity. Three resistance-determining regions related to macrolide and fluoroquinolone were sequenced in 154 M. genitalium positive samples, among which 90.3% harbored mutations related to macrolide or fluroquinolone resistance and 67.5% were multidrug-resistant M. genitalium. A2072G in 23S rRNA and Ser83Ile in parC were the most common mutations. M. hominis was associated with manifestations of bacterial vaginosis in female and epididymitis in male. Conclusions: Single or divorced individuals, those with lower education level and individuals without insurance are higher-risk key populations for STIs. The prevalence of antimicrobial-resistant M. genitalium in Shenzhen is high. Detection of M. hominis increased significantly with lower education level and no health insurance coverage, and it is associated with bacterial vaginosis or epididymitis, indicating that M. hominis deserves further attention.
Assuntos
Antibacterianos , Mycoplasma genitalium , Infecções Sexualmente Transmissíveis , Humanos , Estudos Transversais , China/epidemiologia , Feminino , Masculino , Adulto , Mycoplasma genitalium/genética , Mycoplasma genitalium/isolamento & purificação , Mycoplasma genitalium/efeitos dos fármacos , Infecções Sexualmente Transmissíveis/epidemiologia , Infecções Sexualmente Transmissíveis/microbiologia , Adulto Jovem , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Pessoa de Meia-Idade , Farmacorresistência Bacteriana , Adolescente , Fatores de Risco , Fatores Socioeconômicos , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/microbiologia , Testes de Sensibilidade Microbiana , PrevalênciaRESUMO
Background Increasing rates of macrolide and fluroquinolone resistance in Mycoplasma genitalium (MG) are being reported worldwide with resultant treatment failure. Aim We aimed to determine the level of antibiotic resistance of MG in men who have sex with men (MSM) attending a sexually transmitted infections (STIs) clinic in New Delhi, India. Methods Real-time polymerase chain reaction (PCR) assays targeting MgPa and pdhD genes were performed to detect MG rectal, urogenital or oropharyngeal infections in 180 MSM between January 2022 and June 2023. Macrolide resistance-associated mutations (MRM) and quinolone resistance-associated mutations (QRM) were detected by specific amplification of domain V of 23SrRNA gene and appropriate regions of parC and gyrA genes respectively followed by sequencing. PCR-based screening for Chlamydia trachomatis (CT) infection was also performed. Results A total of 13 (7.2%) MSM were positive for MG infection. The most common site of infection was anorectum (8/13; 61.5%) followed by the urethra (5/13; 38.5%). None of the patients had infection at both the sites, and no oropharyngeal MG infection was detected. CT infection was detected in 37 (20.6%) MSM. Of the 13 MG-infected MSM, 6 (46.2%) were co-infected with CT. MRM and QRM were found in five (46.2%) and two (15.4%) strains, respectively. Both Quinolone resistance mutation (QRM)-harbouring strains also harboured MRM. All the five MG isolates carried the MRM A2071G. Both the QRM isolates co-harboured the parC and gyrA single-nucleotide polymorphisms. There was no correlation between the presence of antibiotic resistance and co-infection with CT (P = 0.52). Limitation Because all patients in the study were MSM, the high rate of resistance to macrolides and fluoroquinolones could not be extrapolated for non-MSM patients. Conclusion This is a report of an initial survey of antibiotic resistance to MG in a country where its diagnosis and treatment are not routinely available. We found a high prevalence of MG-carrying MRM, QRM and dual-class resistance in MSM in the absence of antibiotic exposure. This study mandates the need for both screening and detection of antimicrobial resistance against MG.
Assuntos
Antibacterianos , Farmacorresistência Bacteriana , Fluoroquinolonas , Macrolídeos , Mutação , Infecções por Mycoplasma , Mycoplasma genitalium , Humanos , Mycoplasma genitalium/genética , Mycoplasma genitalium/efeitos dos fármacos , Masculino , Fluoroquinolonas/farmacologia , Fluoroquinolonas/uso terapêutico , Projetos Piloto , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/tratamento farmacológico , Infecções por Mycoplasma/microbiologia , Índia/epidemiologia , Adulto , Farmacorresistência Bacteriana/genética , Macrolídeos/farmacologia , Macrolídeos/uso terapêutico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Homossexualidade Masculina , Adulto Jovem , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Mycoplasma genitalium is an emerging etiology of sexually transmitted infections (STIs) with increasing resistance to antimicrobials. Surveillance on the epidemiology of M. genitalium infection and antimicrobial resistance is warranted. METHODS: Between September 2021 and August 2023, people with HIV (PWH) and people without HIV (PWoH) at risk of STIs were screened for M. genitalium infection using a multiplex polymerase-chain-reaction assay of specimens collected from the rectum, urethra, oral cavity, and vagina. The prevalences of resistance-associated mutations (RAMs) of M. genitalium to fluoroquinolones, macrolides, and tetracycline were investigated. RESULTS: During the 2-year study period, 1021 participants were enrolled, including 531 PWH and 490 PWoH. Overall, 83 (8.1%) and 34 (7.6%) participants had M. genitalium infection at baseline and during follow-up, respectively, with the rectum being the most common site of detection (61.5%). With the first course of antimicrobial treatment, 27 of 63 (42.9%) participants with M. genitalium infection were cured during follow-up, including 24 of 58 (41.4%) who received doxycycline monotherapy. The prevalence of RAMs to macrolides, fluoroquinolones, and tetracyclines at baseline were 24.3%, 22.4%, and 7.9%, respectively. Though PWH had more M. genitalium infection (10.2% vs 5.9%, p = 0.01), a higher rate of RAMs to macrolides (41.0% vs 14.7%, p < 0.01) was found in PWoH. CONCLUSIONS: Among high-risk populations, the prevalence of M. genitalium infection was 8.1%. The overall genotypic resistance of M. genitalium to macrolides and fluoroquinolones was moderately high in Taiwan. Detection of M. genitalium infection and antimicrobial resistance is warranted to ensure resistance-guided antimicrobial treatments to be administered.
Assuntos
Antibacterianos , Farmacorresistência Bacteriana , Fluoroquinolonas , Macrolídeos , Mutação , Infecções por Mycoplasma , Mycoplasma genitalium , Humanos , Mycoplasma genitalium/genética , Mycoplasma genitalium/efeitos dos fármacos , Mycoplasma genitalium/isolamento & purificação , Macrolídeos/farmacologia , Macrolídeos/uso terapêutico , Taiwan/epidemiologia , Fluoroquinolonas/farmacologia , Fluoroquinolonas/uso terapêutico , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/tratamento farmacológico , Infecções por Mycoplasma/microbiologia , Feminino , Masculino , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Adulto , Farmacorresistência Bacteriana/genética , Prevalência , Pessoa de Meia-Idade , Infecções por HIV/tratamento farmacológico , Infecções por HIV/epidemiologia , Testes de Sensibilidade MicrobianaRESUMO
A laboratory-developed test (LDT) using analyte-specific reagents has been optimized on a commercial platform to detect macrolide resistance-associated mutations (MRM) in 23S rRNA from Mycoplasmoides genitalium from primary clinical specimens. In this study, MRM-LDT was applied to a multi-specimen source study set. One thousand four hundred ninety-five primary specimens testing positive for M. genitalium by commercial transcription-mediated amplification (TMA) were initially titered by the TMA assay using serial 10-fold dilutions to semi-quantitate target nucleic acid burden. Primary specimens were then processed for MRM detection using the MRM-LDT. Findings were stratified by gender and specimen source. The mean log10 target nucleic acid titer of a TMA-positive specimen was 3.51 (median 3; range 0-10). Male specimens (n = 1145) demonstrated a mean log10 M. genitalium TMA titer of 3.67; that value observed in 350 female specimens was 2.98 (P < 0.0001). The MRM-LDT detection rate (88.7%) from specimens with log10 M. genitalium TMA titers ≥ 4 was increased over specimens with log10 titers ≤ 1 (4.5%; P < 0.0002). In females, MRM-LDT was positive from 51.3% of vaginal swab and 34.7% of urine specimens (P = 0.01). In males, MRM-LDT was positive from 65.0% of rectal swab and 55.7% of urine specimens (P = 0.002). Differences were also observed in log10 M. genitalium TMA titers as a function of specimen source. M. genitalium macrolide resistance rates among multiple specimen sources, as determined by MRM-LDT, are high in the United States and can be consistent with target nucleic acid burden within the primary specimen. Caveats experienced within subgroupings support MRM reflex testing on primary M. genitalium-positive specimens. IMPORTANCE: First-line macrolide treatment failure is of increasing concern with Mycoplasmoides genitalium in multiple settings. Recent sexually-transmitted infection treatment guidelines from the United States Centers for Disease Control and Prevention have predicated therapeutic approaches on the availability of a macrolide resistance/susceptibility result from a primary clinical specimen. In this report, we investigate potential correlation between macrolide resistance mutation detection rates (identified by a molecular amplified laboratory-developed test) and transcription-mediated amplification-based rRNA target semi-quantitation. Data reveal that rRNA semi-quantitation and laboratory-developed test detection rate differences exist as a function of gender and specimen source. These data can guide providers in proper specimen selection not only for the laboratory diagnosis of M. genitalium but also macrolide resistance mutation determination from primary clinical specimens.
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Farmacorresistência Bacteriana , Macrolídeos , RNA Ribossômico 23S , Humanos , Feminino , Masculino , Macrolídeos/farmacologia , RNA Ribossômico 23S/genética , Farmacorresistência Bacteriana/genética , Fatores Sexuais , Antibacterianos/farmacologia , Mycoplasma genitalium/genética , Mycoplasma genitalium/efeitos dos fármacos , Técnicas de Diagnóstico Molecular/métodos , MutaçãoRESUMO
Sexually transmitted diseases (STDs) are a global concern because approximately 1 million new cases emerge daily. Most STDs are curable, but if left untreated, they can cause severe long-term health implications, including infertility and even death. Therefore, a test enabling rapid and accurate screening and genotyping of STD pathogens is highly awaited. Herein, we present the development of the DNA-based 6STD Genotyping 9G Membrane test, a lateral flow strip membrane assay, for the detection and genotyping of six STD pathogens, including Trichomonas vaginalis, Ureaplasma urealyticum, Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma hominis, and Mycoplasma genitalium. Here, we developed a multiplex PCR primer set that allows PCR amplification of genomic materials for these six STD pathogens. We also developed the six ssDNA probes that allow highly efficient detection of the six STD pathogens. The 6STD Genotyping 9G Membrane test lets us obtain the final detection and genotyping results in less than 30 m after PCR at 25 °C. The accuracy of the 6STD Genotyping 9G membrane test in STD genotyping was confirmed by its 100% concordance with the sequencing results of 120 clinical samples. Therefore, the 6STD Genotyping 9G Membrane test emerges as a promising diagnostic tool for precise STD genotyping, facilitating informed decision-making in clinical practice.
Assuntos
Chlamydia trachomatis , Genótipo , Neisseria gonorrhoeae , Infecções Sexualmente Transmissíveis , Humanos , Chlamydia trachomatis/genética , Chlamydia trachomatis/isolamento & purificação , Neisseria gonorrhoeae/genética , Neisseria gonorrhoeae/isolamento & purificação , Infecções Sexualmente Transmissíveis/microbiologia , Infecções Sexualmente Transmissíveis/diagnóstico , Trichomonas vaginalis/genética , Trichomonas vaginalis/isolamento & purificação , Técnicas de Genotipagem , Mycoplasma hominis/isolamento & purificação , Mycoplasma hominis/genética , Ureaplasma urealyticum/genética , Ureaplasma urealyticum/isolamento & purificação , DNA , Mycoplasma genitalium/genética , Mycoplasma genitalium/isolamento & purificação , Técnicas Biossensoriais , DNA Bacteriano/análise , Reação em Cadeia da Polimerase Multiplex/métodosRESUMO
Sexually transmitted infections (STI) are a public health problem. Real-time PCR assays are the most sensitive test for screening and diagnosis of these infections. The aim of this study was to evaluate a new CT/NG/TV/MG Real-Time PCR (RT-PCR) kit (Vircell) for the detection of Chamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium and Trichomonas vaginalis for the diagnosis of sexual transmitted infections using the Allplex STI Essential Assay (Seegene) as the reference's method. A total of 497 samples from different anatomical sites (endocervical, urethral, rectal, pharyngeal and urine) were analysed from October 2022 to February 2023. A total of 108 (21.73â%) and 106 (21.33â%) positive samples were found for any of the assays used. The most commonly detected pathogen was N. gonorrhoeae (52 samples; 10.46â%), and the least commonly detected was T. vaginalis (three samples; 0.60â%). The anatomical site with the highest prevalence of micro-organisms was a non-urogenital site, the pharynx (26 positive samples; 5.23â%). Using the Allplex STI Essential Assay (Seegene) as the reference method, the diagnosis performance showed that the average specificity of CT/NG/TV/MG RT-PCR Kit (Vircell) was 99.84â% and the sensitivity was 99.53â%. The overall concordance was k=0.98 (CI95â%; 0.96-1). In conclusion, the CT/NG/TV/MG RT-PCR Kit (Vircell) assay shows a good sensitivity and specificity and constitutes a promising and additional alternative to routine procedures for distinct types of clinical specimen in diagnosis STI.
Assuntos
Infecções por Chlamydia , Gonorreia , Infecções por Mycoplasma , Mycoplasma genitalium , Infecções Sexualmente Transmissíveis , Trichomonas vaginalis , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Chlamydia trachomatis/genética , Infecções Sexualmente Transmissíveis/diagnóstico , Infecções Sexualmente Transmissíveis/epidemiologia , Trichomonas vaginalis/genética , Neisseria gonorrhoeae/genética , Mycoplasma genitalium/genética , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/epidemiologia , Tomografia Computadorizada por Raios X , Infecções por Chlamydia/diagnóstico , Gonorreia/diagnóstico , Gonorreia/epidemiologiaRESUMO
Background: Mycoplasmoides genitalium remains a difficult sexually-transmitted infection (STI) to manage due to its potential for antimicrobial resistance and post-infection sequelae. University students are especially vulnerable, as this demographic has the highest rate of STI in the United States. As a result, investigating prevalence rates and therapeutic outcomes in this population is essential to minimize future impact of M. genitalium The purpose of this study was to investigate a university student population for M. genitalium distribution and treatment outcome.Design: Retrospective chart-review of university health clinic attendees, augmented by laboratory detection of M. genitalium following therapeutic intervention.Methods: A total of 1617 student encounters at a midwestern United States university health clinic over a 28-month interval from November 2017 through February 2020 were analyzed for M. genitalium and Chlamydia trachomatis positivity rates and prevalence. Detection of these sexually-transmitted pathogens occurred by commercial RNA amplification testing. Chart review was focused on participant outcomes following initial M. genitalium detection and therapeutic intervention.Results: C. trachomatis positivity and prevalence rates were 7.05% and 9.00%, respectively, while analogous rates for M. genitalium were 7.05% and 6.51%, respectively. An average of 1.83 positive results was generated from participants infected with M. genitalium at any time, with an average of 1.17 positive results for C. trachomatis (P < 0.0002). For students treated with azithromycin, 30.3% generated a negative M. genitalium result upon follow-up, with 1g daily and 2-day 500mg dosing regimens demonstrating less efficacy than a 4-day 250mg regimen or moxifloxacin.Conclusion: Data indicate a need for molecular M. genitalium macrolide resistance determination from primary specimens in the university setting.
Assuntos
Antibacterianos , Mycoplasma genitalium , Humanos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana , Macrolídeos/uso terapêutico , Estudos Retrospectivos , Universidades , Chlamydia trachomatis , Mycoplasma genitalium/genéticaRESUMO
OBJECTIVE: Mycoplasma genitalium (MG) is a microorganism related to sexually transmitted infections. Antibiotic resistance of MG leads to an increase in treatment failure rates and the persistence of the infection. The aim of this study was to describe the most frequent mutations associated with azithromycin and moxifloxacin resistance in our geographical area. METHODS: A prospective study from May 2019 to May 2023 was performed. MG-positive samples were collected. Real-time PCRs (AllplexTM MG-AziR Assay and AllplexTM MG-MoxiR Assay, Seegene) were performed in MG positive samples to detect mutations in 23S rRNA V domain and parC gene. RESULTS: A 37.1% of samples presented resistance determinants to azithromycin and the most common mutation detected was A2059G (57.9%). Resistance to moxifloxacin was studied in 72 azithromycin-resistant samples and 36.1% showed mutations, being G248T the most prevalent (73.1%). CONCLUSIONS: The resistance to different lines of treat ment suggests the need for a targeted therapy and the performing of a test of cure afterwards.
Assuntos
Antibacterianos , Azitromicina , Farmacorresistência Bacteriana , Moxifloxacina , Mutação , Infecções por Mycoplasma , Mycoplasma genitalium , Mycoplasma genitalium/efeitos dos fármacos , Mycoplasma genitalium/genética , Moxifloxacina/farmacologia , Moxifloxacina/uso terapêutico , Azitromicina/farmacologia , Azitromicina/uso terapêutico , Espanha , Humanos , Estudos Prospectivos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana/genética , Infecções por Mycoplasma/tratamento farmacológico , Infecções por Mycoplasma/microbiologia , Feminino , Masculino , Testes de Sensibilidade Microbiana , RNA Ribossômico 23S/genética , Adulto , DNA Topoisomerase IV/genéticaRESUMO
Mycoplasma genitalium (M. genitalium) poses a significant public health challenge due to its association with non-gonococcal urethritis (particularly in men) and antimicrobial resistance. However, despite the prevalence of M. genitalium infections and the rise in resistance rates, routine testing and surveillance remain limited. This is the first study from Croatia that aimed to assess the prevalence and trends of resistance in M. genitalium strains isolated from male individuals by detecting macrolide and fluoroquinolone resistance genes. The study also aimed to explore the factors associated with resistance and changes in resistance patterns over time. Urine samples collected from male individuals in the Zagreb County and northwest region of Croatia between 2018 and 2023 were tested for M. genitalium with the use of molecular methods. Positive samples were subjected to DNA extraction and multiplex tandem polymerase chain reaction (MT-PCR) targeting genetic mutations associated with macrolide (23S rRNA gene) and fluoroquinolone (parC gene) resistance. Of the 8073 urine samples tested from 6480 male individuals (and following the exclusion of repeated specimens), we found that the prevalence of M. genitalium infection was 2.2%. Macrolide resistance was observed in 60.4% of strains, while fluoroquinolone resistance was found in 19.2%. Co-resistance to both antibiotics was present in 18.2% of cases. A statistically significant increase in fluoroquinolone resistance was noted over the study period (p = 0.010), but this was not evident for azithromycin resistance (p = 0.165). There were no statistically significant differences in resistance patterns between age groups, whereas re-testing of patients revealed dynamic changes in resistance profiles over time. The high burden of macrolide resistance and increasing fluoroquinolone resistance underscore the urgent need for comprehensive resistance testing and surveillance programs. The implementation of resistance-guided treatment strategies, along with enhanced access to molecular diagnostics, is pivotal for effectively managing M. genitalium infections.
Assuntos
Antibacterianos , Farmacorresistência Bacteriana , Fluoroquinolonas , Macrolídeos , Infecções por Mycoplasma , Mycoplasma genitalium , Mycoplasma genitalium/genética , Mycoplasma genitalium/efeitos dos fármacos , Mycoplasma genitalium/isolamento & purificação , Humanos , Masculino , Fluoroquinolonas/farmacologia , Fluoroquinolonas/uso terapêutico , Croácia/epidemiologia , Macrolídeos/farmacologia , Macrolídeos/uso terapêutico , Adulto , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/tratamento farmacológico , Infecções por Mycoplasma/urina , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana/genética , Pessoa de Meia-Idade , Adulto Jovem , RNA Ribossômico 23S/genética , Adolescente , Uretrite/microbiologia , Uretrite/epidemiologia , Uretrite/tratamento farmacológico , Testes de Sensibilidade MicrobianaAssuntos
Reação em Cadeia da Polimerase Multiplex , Mycoplasma genitalium , Centros de Atenção Terciária , Humanos , Portugal , Feminino , Mycoplasma genitalium/genética , Mycoplasma genitalium/isolamento & purificação , Infecções por Mycoplasma/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Masculino , Adulto , Fatores de Tempo , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) are routinely tested and reported; however, Trichomonas vaginalis (TV) is the most common sexually transmitted infection (STI) in the United States and the prevalence of Mycoplasma genitalium (MG) infections is likely higher than estimated. We examined the clinical performance of the Alinity m STI assay for detection and surveillance of CT/NG/TV/MG in urine specimens from patients at a large academic medical center. METHODS: Urine specimen from 198 patients was tested in this evaluation. Alinity m STI and Aptima Combo 2 CT/NG and TV assay (Panther System) results were compared, with discrepant results run on the cobas 6800 CT/NG, TV/MG assays. Analyzer turnaround times, time from loading the specimen on the analyzer to results reporting, were determined for Alinity m and Panther systems. RESULTS: Overall percent agreements of the Alinity m in comparison with the Aptima and cobas assays for CT, NG, TV, and MG were 99.5% (97.2%, 99.9%), 99.5% (97.2%, 99.9%), 98.4% (95.5%, 99.5%), and 86.4% (66.7%, 95.3), respectively. There were 5 discrepant samples (CT, 1; NG, 1; TV, 3) between the Alinity m and the Aptima assays, and 3 MG discrepant samples between the Alinity m STI and cobas 6800. Two of the 5 Aptima and Alinity m discrepant samples were resolved as they yielded similar results on both Alinity m and cobas 6800. TV and MG infections comprised 54% of the positive samples and were more often asymptomatic than CT and NG infections. Analyzer turnaround time was 3 hours 25 minutes for the Aptima CT/NG, 3 hours 25 minutes for Aptima TV, and 1 hour 55 minutes for Alinity m STI assay. CONCLUSIONS: The Alinity m STI assay allows for fast and simultaneous detection of the 4 major STI pathogens, which can facilitate surveillance and provide accurate results to help clinicians diagnose for initiation of appropriate treatment.
Assuntos
Chlamydia trachomatis , Gonorreia , Reação em Cadeia da Polimerase Multiplex , Mycoplasma genitalium , Neisseria gonorrhoeae , Trichomonas vaginalis , Humanos , Feminino , Chlamydia trachomatis/isolamento & purificação , Chlamydia trachomatis/genética , Reação em Cadeia da Polimerase Multiplex/métodos , Neisseria gonorrhoeae/isolamento & purificação , Neisseria gonorrhoeae/genética , Trichomonas vaginalis/isolamento & purificação , Trichomonas vaginalis/genética , Masculino , Mycoplasma genitalium/isolamento & purificação , Mycoplasma genitalium/genética , Gonorreia/diagnóstico , Infecções Sexualmente Transmissíveis/diagnóstico , Infecções Sexualmente Transmissíveis/microbiologia , Infecções por Chlamydia/diagnóstico , Infecções por Mycoplasma/diagnóstico , Sensibilidade e Especificidade , Adulto , Estados Unidos , Vaginite por Trichomonas/diagnóstico , Vaginite por Trichomonas/epidemiologia , Kit de Reagentes para DiagnósticoRESUMO
BackgroundAntimicrobial resistance (AMR) of Mycoplasma genitalium (MG) is a growing concern worldwide and surveillance is needed. In Belgium, samples are sent to the National Reference Centre of Sexually Transmitted Infections (NRC-STI) on a voluntary basis and representative or robust national AMR data are lacking.AimWe aimed to estimate the occurrence of resistant MG in Belgium.MethodsBetween July and November 2022, frozen remnants of MG-positive samples from 21 Belgian laboratories were analysed at the NRC-STI. Macrolide and fluoroquinolone resistance-associated mutations (RAMs) were assessed using Sanger sequencing of the 23SrRNA and parC gene. Differences in resistance patterns were correlated with surveillance methodology, socio-demographic and behavioural variables via Fisher's exact test and logistic regression analysis.ResultsOf the 244 MG-positive samples received, 232 could be sequenced for macrolide and fluoroquinolone RAMs. Over half of the sequenced samples (55.2%) were resistant to macrolides. All sequenced samples from men who have sex with men (MSM) (24/24) were macrolide-resistant. Fluoroquinolone RAMs were found in 25.9% of the samples and occurrence did not differ between socio-demographic and sexual behaviour characteristics.ConclusionAlthough limited in sample size, our data suggest no additional benefit of testing MG retrieved from MSM for macrolide resistance in Belgium, when making treatment decisions. The lower occurrence of macrolide resistance in other population groups, combined with emergence of fluoroquinolone RAMs support macrolide-resistance testing in these groups. Continued surveillance of resistance in MG in different population groups will be crucial to confirm our findings and to guide national testing and treatment strategies.
Assuntos
Infecções por Mycoplasma , Mycoplasma genitalium , Minorias Sexuais e de Gênero , Infecções Sexualmente Transmissíveis , Masculino , Humanos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Homossexualidade Masculina , Mycoplasma genitalium/genética , Bélgica/epidemiologia , Macrolídeos/farmacologia , Farmacorresistência Bacteriana/genética , Infecções por Mycoplasma/tratamento farmacológico , Infecções por Mycoplasma/epidemiologia , Mutação , RNA Ribossômico 23S/genética , Fluoroquinolonas/farmacologiaAssuntos
Reação em Cadeia da Polimerase Multiplex , Mycoplasma genitalium , Centros de Atenção Terciária , Humanos , Portugal , Feminino , Mycoplasma genitalium/genética , Mycoplasma genitalium/isolamento & purificação , Infecções por Mycoplasma/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Masculino , Adulto , Fatores de Tempo , Pessoa de Meia-IdadeAssuntos
Antibacterianos , Farmacorresistência Bacteriana , Infecções por Mycoplasma , Humanos , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/tratamento farmacológico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Feminino , Mycoplasma genitalium/efeitos dos fármacos , Mycoplasma genitalium/genética , Mycoplasma genitalium/isolamento & purificação , Testes de Sensibilidade Microbiana , MasculinoRESUMO
BACKGROUND: Mycoplasma genitalium is a sexually transmitted infection (STI) increasing in prevalence. The recent availability of nucleic acid amplification tests (NAATs) has led to updated diagnostic and treatment guidelines. As medication therapy experts, pharmacists can facilitate appropriate antimicrobial selection and stewardship and optimize best patient-care practices in the setting of M. genitalium infection. OBJECTIVE: This study aimed to evaluate patient demographics, therapeutic approaches, and complications of patients with laboratory evidence of M. genitalium hypothesizing that younger adolescent females are affected by this organism, receive suboptimal treatment, and have more complications than adults. METHODS: This was a retrospective cohort study using TriNetX multicenter electronic health record data of subjects aged 12 years and older with evidence of M. genitalium DNA detected via NAATs. The cohort was divided into 2 age groups: adolescents (12-21 years) and adults (older than 21 years). We evaluated age, sex, race, ethnicity, diagnostic codes, and medication codes. RESULTS: Our study included 1126 subjects (192 adolescents [17.1%] and 934 adults [82.9%]) who tested positive for M. genitalium. Subjects in the adolescent group had higher odds of being women (2.52 [1.80, 3.54], P < 0.001), having inflammatory diseases of female pelvic organs diagnostic codes (1.51 [1.06, 2.16], P = 0.025), increased odds of azithromycin prescription (1.70 [1.17, 2.48], P = 0.005), and decreased odds of moxifloxacin prescription (0.41 [0.26, 0.64], P < 0.001). CONCLUSIONS: Our study revealed a higher prevalence of M. genitalium infection in adults and adolescents with increased odds of receiving azithromycin and decreased odds of receiving moxifloxacin. Both age groups had decreased odds of receiving doxycycline compared with azithromycin despite guidelines recommending initial empirical antibiotic treatment with doxycycline and growing macrolide resistance. Suboptimal treatment of this infection may lead to lifelong complications. Pharmacists may provide crucial guidance and education to both patients and health care providers regarding appropriate treatment for M. genitalium.
Assuntos
Infecções por Mycoplasma , Mycoplasma genitalium , Adulto , Adolescente , Humanos , Feminino , Criança , Adulto Jovem , Masculino , Antibacterianos , Azitromicina/uso terapêutico , Azitromicina/farmacologia , Moxifloxacina/uso terapêutico , Moxifloxacina/farmacologia , Estudos Retrospectivos , Doxiciclina/farmacologia , Doxiciclina/uso terapêutico , Mycoplasma genitalium/genética , Registros Eletrônicos de Saúde , Macrolídeos/uso terapêutico , Macrolídeos/farmacologia , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/tratamento farmacológico , Infecções por Mycoplasma/epidemiologia , Farmacorresistência Bacteriana/genética , PrevalênciaAssuntos
Proteínas de Bactérias , DNA Girase , Farmacorresistência Bacteriana , Infecções por Mycoplasma , Mycoplasma genitalium , Humanos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Austrália/epidemiologia , Fluoroquinolonas/farmacologia , Macrolídeos , Mutação , Mycoplasma genitalium/genética , Infecções por Mycoplasma/tratamento farmacológico , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/microbiologia , Queensland , DNA Girase/genética , DNA Girase/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismoRESUMO
Background: Traditional drug susceptibility testing cannot be performed in clinical laboratories due to the slow-growing characteristics of Mycoplasma genitalium when cultured in vitro. Sanger sequencing is the standard method for detecting drug resistance-associated mutations. It has been used in some laboratories to guide the choice of macrolide antibiotics for Mycoplasma genitalium infected patients. Furthermore, resistance to fluoroquinolone has become another emerging clinical challenge. Objective: Sequencing analysis can detect unknown mutations, but it is time-consuming, requires professional analytical skills and the appropriate testing equipment. The main objective of this study was to establish a nested real-time PCR method for the simultaneous detection of 23S rRNA and parC genotypes in relation to the macrolide and fluoroquinolone resistance. Results: 105 MG-positive samples and 27 samples containing other pathogens were used for validation. The limit of the nested real-time PCR detection was 500 copies/reaction and there was no cross-reaction with Ureaplasma urealyticum, Mycoplasma hominis, Chlamydia trachomatis, Neisseria gonorrhoeae, Human papillomavirus, Herpes simplex virus, Candida albicans and Ureaplasma parvum, but the 23S rRNA assay cross-reacted with Mycoplasma pneumoniae. Compared with sequencing results, the sensitivity of 23S rRNA was 100% (95% CI; 93.3 -100), the specificity was 94.3% (95% CI; 79.4 - 99.0), the overall consistency was 98% (95% CI; 92.5 - 99.7) and kappa value was 0.96 (P < 0.001); the sensitivity of parC was 100% (95% CI; 93.4 - 100), the specificity was 89.7% (95% CI; 71.5 - 97.3) and the overall consistency was 96.9% (95% CI; 90.7 - 99.2) with a kappa value of 0.92 (P < 0.001). Conclusions: The results of this sensitive and rapid alternative for identifying resistant genotypes of Mycoplasma genitalium are intuitive and easy to interpret, especially for mixed MG populations. Although the relevant 23S rRNA primers need further adjustment, this reliable method would provide an effective diagnostic tool for the selection of antibiotics in clinical practice.
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Mycobacterium tuberculosis , Infecções por Mycoplasma , Mycoplasma genitalium , Humanos , Fluoroquinolonas/farmacologia , Mycoplasma genitalium/genética , RNA Ribossômico 23S/genética , Reação em Cadeia da Polimerase em Tempo Real , Macrolídeos/farmacologia , Testes de Sensibilidade Microbiana , Farmacorresistência Bacteriana/genética , Infecções por Mycoplasma/diagnóstico , Mycobacterium tuberculosis/genética , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , MutaçãoRESUMO
The LightMix® Modular Mycoplasma Macrolide and LightMix® Modular parC Fluoroquinolone Resistance assays (TIB Molbiol) were evaluated using sequential Mycoplasma genitalium positive (n = 125) and negative (n = 93) clinical samples. Results were compared to the results of an established commercial assay (ResistancePlus MG assay, SpeeDx Pty Ltd) or Sanger sequencing (for parC). Detection of M. genitalium by the TIB Molbiol assay had a high agreement with the reference assay, with a positive percent agreement (PPA) of 97.6 [95% confidence interval (CI): 93.1-99.5] and negative percent agreement (NPA) of 95.7 (95% CI: 89.5-98.8). From 105 positive samples, macrolide resistance detection had a PPA of 100% (95% CI: 93.7-100) and NPA of 81.3% (95% CI: 67.4-91.1). For the detection of fluroquinolone resistance mutation G248T/S83I or "other mutation" in the quinolone resistance determinant region, from 95 samples there was 100% (95% CI: 86.3-100) sensitivity and 100% (95% CI: 94.5-100) specificity. The understanding of the basis for fluoroquinolone treatment failure is still developing; it is therefore important to use the output of parC-based resistance assays with caution to avoid the inappropriate use of antibiotic therapies, especially considering the limited number of alternative treatments.
Assuntos
Infecções por Mycoplasma , Mycoplasma genitalium , Humanos , Antibacterianos/farmacologia , Fluoroquinolonas , Macrolídeos , Mycoplasma genitalium/genética , Farmacorresistência Bacteriana/genética , Infecções por Mycoplasma/diagnóstico , Mutação , RNA Ribossômico 23S/genética , PrevalênciaRESUMO
BACKGROUND: Diagnosis of infected individuals with Mycoplasma genitalium (MG) is often performed by real-time PCR or transcription-mediated amplification (TMA). A limitation of the MG-TMA assay is the relatively short time span of 24 h in which the collected urine is required to be transferred into a Urine Specimen Transport Tube, according to the manufacturer's guidelines. If not transferred within 24 h, the manufacturer's claimed sensitivity cannot be guaranteed anymore, and samples may instead be tested with an in-house validated real-time PCR, despite its recognized lower sensitivity. This study aimed to validate an exception to the sample transport and storage conditions of the MG-TMA assay as set by the manufacturer, being the prolongation of the acceptable testing time limit of 24 h. METHODS: From June to December 2022, first-void urines were collected from clients attending the clinic for sexual health in Amsterdam, the Netherlands. Urine samples that tested positive for MG by TMA assay at the day of collection were concomitantly stored at room (18-24 °C) and refrigerator temperature (4-8 °C) for 15 days. The stored urine samples were tested with both an in-house validated real-time PCR and MG-TMA assay after transfer of the original urine samples to the respective test tubes at 3, 7, 12 and 15 days post collection. RESULTS: In total, 47 MG-positive urine samples were collected, stored and tested for MG by real-time PCR and TMA assays. After storage at room temperature, the MG-detection rate by TMA was significantly higher compared to real-time PCR, at days 0 (p ≤ 0.001), 7 (p ≤ 0.001) and 12 (p < 0.05). After storage at refrigerator temperature, the MG-detection rate determined by TMA assay was significantly enhanced in comparison with real-time PCR at days 3 (p < 0.01), 7 (p ≤ 0.001) and 15 (p < 0.01). CONCLUSIONS: This validation study showed that the MG-TMA assay has a superior detection rate in urine compared to real-time PCR, up to 15 days post sample collection and irrespective of storage temperature. Accepting urines older than 24 h to be tested by TMA will improve clinical diagnosis of MG infections.
Assuntos
Líquidos Corporais , Mycoplasma genitalium , Humanos , Mycoplasma genitalium/genética , Reação em Cadeia da Polimerase em Tempo Real , Instituições de Assistência Ambulatorial , Capsaicina , MentolRESUMO
OBJECTIVE: Macrolide and fluoroquinolone resistance in Mycoplasma genitalium (MG) is of emerging global concern. Compared with neighbouring countries such as Denmark, Sweden has had lower rates of macrolide resistance while fluoroquinolone resistance rates are less well documented. This study retrospectively examined macrolide, fluoroquinolone and multidrug resistance rates from Dalarna County, Sweden over a 13-year period. METHODS: MG-positive samples from 2006 to 2018 from patients examined at the Department of Venereology, Central Hospital, Falun, Sweden were tested by sequencing for macrolide resistance mutations (MRM) and fluoroquinolone resistance-associated mutations (QRAM) in the parC and gyrA subunit regions. A subset of these samples from 2006 to 2011 have been reported on previously, although only for MRM. RESULTS: Of 874 samples, 98 (11.2%, 95% CI 9.1% to 13.6%) had mutations associated with resistance to macrolides and 19 of 828 (2.3%, 95% CI 8.9% to 23.1%) to quinolones. Mutations associated with resistance to both drugs were detected in 5 of 828 (0.6%, 95% CI 0.1% to 1.4%) samples overall. A significant positive linear trend (p=0.004) for an increase in the rate of macrolide resistance was observed (from 0% in 2006 to 31% in 2018) while the increase in QRAM from 0% in 2006 to 12.3% in 2018 was not statistically significant. CONCLUSIONS: Despite a decrease in macrolide and fluoroquinolone consumption in Sweden, there was an overall increase in MG macrolide, fluoroquinolone and dual resistance from 2006 to 2018, although the difference in fluoroquinolone resistance rates was not statistically significant. In order to maintain comparably low resistance rates, resistance-guided therapy for MG infections will be crucial.