In VitroActivity of the New Fluoroketolide Solithromycin (CEM-101) against Macrolide-Resistant and -Susceptible Mycoplasma genitalium Strains

ABSTRACTMycoplasma genitaliumhas become well established as an etiological agent of sexually transmitted infections, but due to its fastidious growth requirements, only a fewM. genitaliumstrains are available to determine the MICs of currently used and new antimicrobial agents. Recent clinical trials have suggested that treatment with azithromycin has decreasing efficacy due to an increasing prevalence of macrolide resistance, and alternative treatment with moxifloxacin is similarly under pressure from emerging resistance. Thus, there is an urgent need for new antimicrobials. Thein vitroactivity of the newly developed fluoroketolide solithromycin (CEM-101) was evaluated against a collection of 40M. genitaliumstrains, including 15 with high-level macrolide resistance and 5 multidrug-resistant strains with resistance to both macrolides and quinolones. Furthermore, the MIC of solithromycin was correlated with mutations in the 23S rRNA gene and in the genes encoding ribosomal proteins L4 and L22. Thein vitroresults showed that solithromycin has activity againstM. genitaliumsuperior to that of other macrolides, doxycycline, and fluoroquinolones. Accordingly, this new fluoroketolide might be an effective option for treatment ofM. genitaliuminfections. However, the efficacy of solithromycin in clinical trials with follow-up for test of cure and detection of genotypic and phenotypic resistance needs to be evaluated prior to widespread use. In a phase 2 clinical trial, solithromycin was highly effective as a single oral dose againstC. trachomatisandNeisseria gonorrhoeae, suggesting that solithromycin could be a treatment option for several sexually transmitted infections, including in syndromic treatment of urethral and vaginal discharge.

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Prevalence of Mycoplasma genitalium and Antibiotic Resistance-Associated Mutations in Patients at a Sexually Transmitted Infection Clinic in Iceland, and Comparison of the S-DiaMGTV and Aptima Mycoplasma genitalium Assays for Diagnosis

Journal of Clinical Microbiology ◽

10.1128/jcm.01084-20 ◽

2020 ◽

Vol 58 (9) ◽

Author(s):

Ingibjörg Hilmarsdóttir ◽

Eva Mjöll Arnardóttir ◽

Elísabet Reykdal Jóhannesdóttir ◽

Freyja Valsdóttir ◽

Daniel Golparian ◽

...

Keyword(s):

Sexually Transmitted Infection ◽

False Negative ◽

Mycoplasma Genitalium ◽

23S Rrna ◽

Rrna Gene ◽

Transmitted Infection ◽

Content Type ◽

Sexually Transmitted ◽

Negative Results

ABSTRACT Mycoplasma genitalium is prevalent among attendees in sexually transmitted infection (STI) clinics, and therapy is hampered by rapidly rising levels of resistance to azithromycin and moxifloxacin. In this study, we evaluated, for the first time in Iceland, the prevalence of M. genitalium and azithromycin and moxifloxacin resistance-associated mutations and assessed the diagnostic performance of the CE/in vitro diagnosis (IVD)-marked S-DiaMGTV (Diagenode Diagnostics) versus the U.S. FDA/CE/IVD-approved Aptima MG (AMG; Hologic) for M. genitalium detection. From October 2018 to January 2019, urine and vaginal swabs were provided by male and female attendees at Iceland’s only STI clinic. Specimens were tested with S-DiaMGTV and AMG, and resistance-associated mutations were determined by 23S rRNA gene and parC sequencing. Demographic and clinical data were collected from patient records. M. genitalium prevalence was 9.3% overall; 7.7% (38/491) among male and 10.9% (53/487) among female participants. Azithromycin and moxifloxacin resistance-associated mutations were found in 57.0% (45/79) and 0.0% (0/80) of evaluable specimens, respectively. Sensitivity was 72.5% and 100%, and specificity was 99.9% and 100% for S-DiaMGTV and AMG, respectively. No association was found between M. genitalium and symptoms of urethritis in men. Prevalence rates for M. genitalium and azithromycin resistance-associated genes in Iceland are among the highest reported in Europe. The significantly higher sensitivity of AMG over that of S-DiaMGTV can have important clinical implications. More information is urgently needed to clarify the significance of false-negative results obtained with S-DiaMGTV and other similarly performing widely used real-time PCR methods for diagnosis and management of this sexually transmitted infection.

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Prospective Evaluation of ResistancePlus MG, a New Multiplex Quantitative PCR Assay for Detection of Mycoplasma genitalium and Macrolide Resistance

Journal of Clinical Microbiology ◽

10.1128/jcm.02312-16 ◽

2017 ◽

Vol 55 (6) ◽

pp. 1915-1919 ◽

Cited By ~ 41

Author(s):

S. N. Tabrizi ◽

J. Su ◽

C. S. Bradshaw ◽

C. K. Fairley ◽

S. Walker ◽

...

Keyword(s):

Quantitative Pcr ◽

Clinical Performance ◽

Simultaneous Detection ◽

Mycoplasma Genitalium ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Resistance Mutations ◽

Content Type ◽

Conventional Culture

ABSTRACT Mycoplasma genitalium is a significant pathogen for which first-line treatment is becoming less effective due to increased resistance to macrolides. As conventional culture and antimicrobial susceptibility testing is not feasible for routine detection of this pathogen, molecular markers such as detection of mutations in the 23S rRNA gene have been described to predict resistance. Recently, a novel multiplex quantitative PCR (qPCR) assay, ResistancePlus MG, has been described for the simultaneous detection of Mycoplasma genitalium and macrolide resistance. In the current study, the clinical performance of the assay was evaluated on 1,089 consecutive urine and anogenital swab samples in symptomatic and asymptomatic male and female patients. Overall, 6.0% were positive for M. genitalium , with 63.1% having macrolide resistance-associated mutations. Compared to the laboratory-validated qPCR method targeting the 16S rRNA gene and Sanger sequencing to determine 23S rRNA mutations, the sensitivity and specificity of M. genitalium detection were 98.5% and 100% and for detection of macrolide resistance mutations were 100.0% and 96.2%, respectively. This assay offers a considerable advantage in clinical settings for M. genitalium testing by making the results of macrolide resistance and mutation analyses simultaneously available, which is increasingly important with escalating macrolide resistance.

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Mycoplasma genitalium in Primary Care: Prevalence and azithromycin resistance in Santiago de Compostela Health Care Area

Revista Española de Quimioterapia ◽

10.37201/req/052.2021 ◽

2021 ◽

Author(s):

Mercedes Treviño ◽

María Rodríguez-Velasco ◽

Tamara Manso ◽

María Cea

Keyword(s):

Primary Care ◽

Health Care ◽

General Population ◽

Sexually Transmitted Infections ◽

Mycoplasma Genitalium ◽

23S Rrna ◽

Rrna Gene ◽

Santiago De Compostela ◽

Sexually Transmitted ◽

Azithromycin Resistance

Objectives. Mycoplasma genitalium is associated with persistent/recurrent sexually transmitted infections. The aim of this work was to estimate the prevalence and azithromycin resistance of M. genitalium in general population that was attended at Primary Care of Santiago de Compostela Health Care Area. Material and methods. The study was carried out in 2019 in general population of Santiago de Compostela Health Care Area. Real-time multiplex PCR was used for screening of sexually transmitted infections associated pathogens and detection of mutations in the 23S rRNA gene. Results. A total of 502 women and 532 men were studied. The prevalence of M. genitalium was 2,4% in men and 2,9% in women. Overall azithromycin resistance was 20% all of them detected in men. The mutations found were A2059G, A2058G and A2058T. Conclusions. Although the proportion of M. genitalium infection is low, the high percentage of azithromycin resistance detected supports the relevance of these data in order to the right management of the patients with sexually transmitted diseases and, so as, to avoid the emergence of resistance in other pathogens of the urogenital tract.

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Macrolide Resistance in Mycoplasma genitalium in Singapore

Open Forum Infectious Diseases ◽

10.1093/ofid/ofx163.093 ◽

2017 ◽

Vol 4 (suppl_1) ◽

pp. S104-S104 ◽

Cited By ~ 2

Author(s):

Timothy Barkham ◽

Wen Ying Tang ◽

Siti Aminah Mansoor ◽

Martin Tze-Wei Chio

Keyword(s):

Direct Detection ◽

Bacterial Genome ◽

Mycoplasma Genitalium ◽

Clinical Diagnostics ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Resistance Mutations ◽

Transmitted Infection ◽

Sexually Transmitted

Abstract Background Mycoplasma genitalium was first reported as a cause of non-gonococcal urethritis in 1980. It has progressed from being an ‘emerging’ sexually transmitted infection (STI) to an accepted STI. Prevalence of infection has been reported as the Netherlands 4.5%, Sweden 6.3%, UK 1.2% and France 4%. M. genitalium has the smallest known bacterial genome and was the second bacterial genome fully sequenced. It has minimal requirements and is said to approach the minimum possible for a living cell. It is extremely fastidious; only a few strains have been cultured worldwide. Diagnosis relies on direct detection. It does not have a cell wall so it is not susceptible to antibiotics such as penicillins and cephalosporins. Therapy depends on fluoroquinolones and macrolides but resistance to macrolides has been widely reported: 13% France, 18% Sweden, 40% UK, Australia and Denmark, 100% Greenland, 30% Japan. Methods Ethics approval was granted. DNA extracts left over after routine clinical diagnostics at the Department of STI Control (DSC) Clinic, Kelantan Lane, Singapore were harvested. DNA had been extracted on a Cobas 4800 instrument (Roche) from urine and urethral swabs collected for testing for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG). A 2-plex real-time PCR assay targeting the pdhD and mgpB genes was used to screen for M. genitalium. Samples were deemed positive if both targets were detected. If only one target was detected, the sample was retested; if reactive in either target upon retest, the sample was considered positive for M. genitalium. Positive DNA preps were then screened for macrolide resistance mutations after Sanger sequencing of the 23S rRNA gene. Results 368 anonymised DNA elutes from 254 urines and 114 urethral swabs were collected between May and July 2016. One hundred eighty-four were CT/NG positive and 184 were CT/NG negative. Sixteen (4.3%) were positive for M. genitalium. Four (25%) of these 16 samples contained macrolide resistance associated mutations; A2058T (x2), A2058G (x1), and A2059G (x1). Conclusion M. genitalium was detected in 4.3% of samples. Macrolide resistance mutations were detected in 25%, similar to international rates. Some guidelines recommend testing for resistance to guide therapy and to perform a test of cure. Disclosures All authors: No reported disclosures.

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Clinical Evaluation of Three Commercial PCR Assays for the Detection of Macrolide Resistance in Mycoplasma genitalium

Journal of Clinical Microbiology ◽

10.1128/jcm.01478-19 ◽

2019 ◽

Vol 58 (2) ◽

Cited By ~ 1

Author(s):

Chloé Le Roy ◽

Cécile Bébéar ◽

Sabine Pereyre

Keyword(s):

Sensitivity And Specificity ◽

Internal Control ◽

Simultaneous Detection ◽

Mycoplasma Genitalium ◽

Macrolide Resistance ◽

University Hospital ◽

23S Rrna ◽

Rrna Gene ◽

Content Type ◽

Commercial Kits

ABSTRACT As macrolide resistance in Mycoplasma genitalium is increasing worldwide, macrolide resistance-associated mutations should be assessed in M. genitalium-positive specimens. New commercial kits are available for detection of macrolide resistance concurrently with M. genitalium. We prospectively evaluated the handling and clinical performances of three commercial kits for detection of macrolide resistance in M. genitalium. Between August and December 2018, remnants of all urogenital specimens determined to be M. genitalium positive using an in-house real-time PCR assay were prospectively collected at the French National Reference Center for Bacterial Sexually Transmitted Infections, Bordeaux University Hospital, Bordeaux, France. The internal control of each kit was added to the primary specimen before DNA extraction, and the absence of amplification inhibition associated with the addition of the three internal controls was assessed. Specimens were evaluated with four assays: the ResistancePlus MG assay (SpeeDx), the S-DiaMGRes assay (Diagenode), the RealAccurate TVMGres assay (PathoFinder), and amplification and sequencing of the 23S rRNA gene (the reference assay). Overall, 195 M. genitalium-positive specimens were assessed. The positive agreement of M. genitalium detection for each kit ranged between 94.8% and 96.4%. Among 154 specimens with M. genitalium positivity as detected by the three commercial kits and 23S rRNA sequencing data, the clinical sensitivity and specificity ranges of the three commercial kits for detecting macrolide resistance-associated mutations were 95 to 100% and 94.6 to 97.3%, respectively. The sensitivity and specificity values were similar among the kits. The launch of three easy-to-use sensitive and specific commercial kits for simultaneous detection of M. genitalium and macrolide resistance will be useful for resistance-guided therapy.

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961. Prevalence and Macrolide Resistance of Mycoplasma genitalium After Initiation of HIV Preexposure Prophylaxis

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz359.063 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S29-S29

Author(s):

Jens Van Praet ◽

Sanne Steyaert ◽

Stefaan Vandecasteele ◽

Barbara Van Den Bergh ◽

Hilde Mahieu ◽

...

Keyword(s):

Mycoplasma Genitalium ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Sexually Transmitted ◽

Preexposure Prophylaxis ◽

23S Rrna Gene ◽

Resistant Strains ◽

Taqman Array

Abstract Background Recent evidence shows that patients using HIV preexposure prophylaxis (PrEP) have an increased rate of bacterial sexually transmitted infections (STIs), including syphilis, chlamydia, and gonorrhea. The rate of Mycoplasma genitalium infections and the susceptibility of M. genitalium in patients on PrEP have been less well described. Methods We studied all patients who started on PrEP in the AZ Sint-Jan Hospital Bruges from January 6, 2017 to January 4, 2019. Patients were screened for M. genitalium and other bacterial STIs with rectal swabs, pharyngeal swabs, first-voided urine and blood collections at baseline and quarterly intervals after initiating PrEP. TaqMan array card technology was used to detect M. genitalium and determine macrolide-resistance mediating mutations in the region V of the 23S rRNA gene (A2058G, A2059G, A2058C, and others). Patients with an STI were treated based on a national guideline. Proportions were estimated using a Generalized Estimating Equations model with independent correlation structure. Results A total of 136 males and 1 female (median age, 40 years (interquartile range (IQR), 20–79)) were included in the study. All men were gay or bisexual. The median follow-up time was 11.3 months (IQR, 4.7–15.3). In total, 117 patients (85%) used PrEP daily at their last visit. The estimated proportion of patients with M. genitalium at baseline, 3 months, 6 months, 9 months, and 12 months was 7% (95% CI 4–13), 12% (95% CI 7–20), 7% (95% CI 4–15), 6% (3–15), and 6% (2–15). Thirty-two patients (23%) tested at least once positive for M. genitalium during the study period. The estimated percentage of macrolide resistance increased from 40% (95% CI 16–70) at baseline to 71% (95% CI 44–89), 67% (95% CI 27–92), 80% (95% CI 31–97), and 75% (95% CI 24–97) at 3 months, 6 months, 9 months, and 12 months, respectively. Conclusion After initiation of PrEP, the prevalence of M. genitalium in our cohort at quarterly screening was not increased compared with baseline. However, a nonsignificant trend of an increased percentage of macrolide-resistant strains was observed. Disclosures All Authors: No reported Disclosures.

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A 5′ Nuclease Genotyping Assay for Identification of Macrolide-Resistant Mycoplasma genitalium in Clinical Specimens

Journal of Clinical Microbiology ◽

10.1128/jcm.00012-16 ◽

2016 ◽

Vol 54 (6) ◽

pp. 1593-1597 ◽

Cited By ~ 15

Author(s):

Gitte Qvist Kristiansen ◽

Jan Gorm Lisby ◽

Kristian Schønning

Keyword(s):

Mycoplasma Genitalium ◽

Antimicrobial Treatment ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Predictive Values ◽

Content Type ◽

Clinical Specimens ◽

Genotyping Assay ◽

23S Rrna Gene

Rapid and sensitive detection of macrolide resistance inMycoplasma genitaliumis required for the guidance of adequate antimicrobial treatment. Previous studies have confirmed that single-base mutations at position 2058 or 2059 in domain V of the 23S rRNA gene ofM. genitaliumresult in high-level macrolide resistance. Sequencing of PCR products remains the gold standard for the identification of mutations conferring resistance to macrolides but is laborious and time-consuming. The aim of the present study was to develop a 5′ nuclease genotyping assay to detect single nucleotide polymorphisms in the 23S rRNA gene ofMycoplasma genitaliumthat are associated with macrolide resistance by combining PCR with hydrolysis probes and subsequent endpoint genotyping analysis. The 5′ nuclease genotyping assay was used as a referral test to be used onM. genitalium-positive samples and was validated on 259 positive samples, of which 253 (97.7%) were successfully sequenced. With the newly developed assay, 237/259 (91.5%) investigatedM. genitalium-positive samples were genotyped. The positive and the negative predictive values were 100% when evaluated on successfully genotyped samples. The newly developed assay discriminated macrolide-resistantM. genitaliumin clinical specimens possessing A2058G, A2058C, A2058T, and A2059G mutations with a sensitivity of 94.4% (95% confidence interval [CI], 90.7% to 98.2%) and a specificity of 92.7% (95% CI, 87.8% to 97.6%) when evaluated on successfully sequenced samples. The assay can correctly guide antimicrobial treatment ofM. genitaliuminfections.

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In Vitro Activity of Lefamulin against Sexually Transmitted Bacterial Pathogens

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02380-17 ◽

2018 ◽

Vol 62 (5) ◽

Cited By ~ 20

Author(s):

Susanne Paukner ◽

Astrid Gruss ◽

Jørgen Skov Jensen

Keyword(s):

Chlamydia Trachomatis ◽

Bacterial Pathogens ◽

Mycoplasma Genitalium ◽

Standard Of Care ◽

Multidrug Resistant ◽

In Vitro Activity ◽

First Line ◽

Content Type ◽

Sexually Transmitted

ABSTRACT The pleuromutilin antibiotic lefamulin demonstrated in vitro activity against the most relevant bacterial pathogens causing sexually transmitted infections (STI), including Chlamydia trachomatis (MIC 50/90 , 0.02/0.04 mg/liter; n = 15), susceptible and multidrug-resistant Mycoplasma genitalium (MIC range, 0.002 to 0.063 mg/liter; n = 6), and susceptible and resistant Neisseria gonorrhoeae (MIC 50/90 , 0.12/0.5 mg/liter; n = 25). The results suggest that lefamulin could be a promising first-line antibiotic for the treatment of STI, particularly in populations with high rates of resistance to standard-of-care antibiotics.

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Levels ofMycoplasma genitaliumAntimicrobial Resistance Differ by Both Region and Gender in the State of Queensland, Australia: Implications for Treatment Guidelines

Journal of Clinical Microbiology ◽

10.1128/jcm.01555-18 ◽

2019 ◽

Vol 57 (3) ◽

Cited By ~ 15

Author(s):

E. L. Sweeney ◽

E. Trembizki ◽

C. Bletchly ◽

C. S. Bradshaw ◽

A. Menon ◽

...

Keyword(s):

Antibiotic Resistance ◽

Treatment Guidelines ◽

Mycoplasma Genitalium ◽

Macrolide Resistance ◽

Quinolone Resistance ◽

Resistance Mutations ◽

Transmitted Infection ◽

Content Type ◽

Sexually Transmitted ◽

And Gender

ABSTRACTMycoplasma genitaliumis frequently associated with urogenital and rectal infections, with the number of cases of macrolide-resistant and quinolone-resistantM. genitaliuminfection continuing to increase. In this study, we examined the levels of resistance to these two common antibiotic treatments in geographically distinct locations in Queensland, Australia. Samples were screened for macrolide resistance-associated mutations using a commercially available kit (ResistancePlus MG; SpeeDx), and quinolone resistance-associated mutations were identified by PCR and DNA sequencing. Comparisons between antibiotic resistance mutations and location/gender were performed. The levels ofM. genitaliummacrolide resistance were high across both locations (62%). Quinolone resistance mutations were found in ∼10% of all samples, with a number of samples harboring mutations conferring resistance to both macrolides and quinolones. Quinolone resistance was higher in southeast Queensland than in north Queensland, and this was consistent in both males and females (P = 0.007). TheM. genitaliumisolates in rectal swab samples from males harbored high levels of macrolide (75.9%) and quinolone (19%) resistance, with 15.5% harboring resistance to both classes of antibiotics. Overall, the lowest observed level of resistance was to quinolones in females from north Queensland (1.6%). These data highlight the high levels of antibiotic resistance inM. genitaliumisolates within Queensland and the challenges faced by sexually transmitted infection clinicians in managing these infections. The data do, however, show that the levels of antibiotic resistance may differ between populations within the same state, which has implications for clinical management and treatment guidelines. These findings also support the need for ongoing antibiotic resistance surveillance and tailored treatment.

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Mycoplasma genitalium in Symptomatic Male Urethritis: Macrolide Use Is Associated With Increased Resistance

Clinical Infectious Diseases ◽

10.1093/cid/ciz294 ◽

2019 ◽

Vol 70 (5) ◽

pp. 805-810 ◽

Cited By ~ 8

Author(s):

Yang Li ◽

Xiaohong Su ◽

Wenjing Le ◽

Sai Li ◽

Zhaoyan Yang ◽

...

Keyword(s):

Ribosomal Rna ◽

Mycoplasma Genitalium ◽

Macrolide Resistance ◽

Rrna Genes ◽

Fluoroquinolone Resistance ◽

23S Rrna ◽

Transmitted Infection ◽

Sexually Transmitted ◽

23S Ribosomal Rna ◽

The Impact

Abstract Background Mycoplasma genitalium (MG) causes symptomatic urethritis in men, and can infect alone or together with other sexually transmitted infection (STI) agents. Methods The prevalence of MG and other STIs was determined in 1816 men with symptomatic urethritis. Resistance of MG to macrolides and fluoroquinolones was determined by sequencing; the impact of recent antimicrobial usage on the distribution of MG single or mixed infections was determined. Results Overall, prevalence of MG infection was 19.7% (358/1816). Fifty-four percent (166/307) of MG infections occurred alone in the absence of other STI agents. Men with single MG infection self-administered or were prescribed antibiotics more often in the 30 days prior to enrollment than subjects with urethritis caused by MG coinfection (P < .0001). Higher rates (96.7%) of infection with macrolide resistance in MG were identified in men who had taken macrolides prior to enrollment (P < .03). Overall, 88.9% (303/341) of 23S ribosomal RNA (rRNA) genes contained mutations responsible for macrolide resistance; 89.5% (308/344) of parC and 12.4% (42/339) of gyrA genes had mutations responsible for fluoroquinolone resistance. Approximately 88% (270/308) of MG had combined mutations in 23S rRNA and parC genes; 10.4% (32/308) had mutations in all 3 genes. Conclusions MG was the single pathogen identified in 11% of men with symptomatic urethritis. Overall, nearly 90% of MG infections were resistant to macrolides and fluoroquinolones. Men who took macrolides in the 30 days prior to enrollment had higher rates (97%) of macrolide-resistant MG. Resistance was associated with numerous mutations in 23SrRNA, parC, and gyrA genes.

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