Persistent and recurrent urethritis due to macrolide-resistant Mycoplasma genitalium: first reports in Argentina

2021 ◽  
Author(s):  
Gabriela Baldoni ◽  
Gabriela Iribarren ◽  
Claudia Garbasz ◽  
Pablo Striebeck ◽  
Micaela Mayer Wolf ◽  
...  
Keyword(s):  
Male Patient ◽  
Mycoplasma Genitalium ◽  
Purulent Discharge ◽  
23S Rrna ◽  
Rrna Gene ◽  
Reference Laboratory ◽  
Sequencing Analysis ◽  
Adequate Treatment ◽  
23S Rrna Gene ◽  
Clinical Cases

Introduction: Mycoplasma genitalium (MG) is responsible for 15%-20% nongonococcal urethritis in men. In Argentina, the diagnosis is only performed by few laboratories. Single-dose 1 g azithromycin (AZM1D) treatment leads to emergence of macrolide resistance (mutations at 23S rRNA gene, region V, position 2058 or 2059). Recommendations include 5-day AZM (AZM5D) regimen, moxifloxacin as second-line therapy. Doxycycline is only 30% effective. Test of Cure (ToC) is advisable. Objective: The aim of this study was to describe the first two clinical cases of persistent and recurrent urethritis due to macrolide-resistant MG in Argentina. Methods: End point polymerase chain reaction (PCR) for diagnosis and ToC. Sanger sequencing analysis of mutations. Results: Case 1: A 26-year-old male patient with occasional heterosexual contacts and no history of sexually transmitted infections (STIs) complained urethral thick purulent discharge and dysuria (January 2018), with negative microbiological cultures and Chlamydia trachomatis PCR. The patient received ceftriaxone/AZM1D. However, symptoms persisted (April 2018). Later, doxycycline was prescribed for 1 month. Five days after treatment, the sample was referred to the STI national reference laboratory (NRL) and results were found positive for MG. The patient was given AZM5D. As a result, symptoms disappeared, posterior ToC was found negative, and retrospectively, sequencing 23S rRNA gene showed A2058G transition. Case 2: An 18-year-old male patient with stable heterosexual relationship complained of previous gonococcal urethritis and urethral serous exudate with inflammatory reaction (September 2017), with negative microbiological cultures. The patient received ceftriaxone and AZM1D as initial treatment. Later, he was given doxycycline for 10 days. On February 2018, symptoms reappeared and sample referred to the NRL was positive for MG (negative for other STIs). With AZM1D treatment, symptoms disappeared. After 1 month, the symptoms recurred. Results showed a new MG-positive sample (April 2018). AZM5D administration induced 2 weeks symptoms free and recurrence, requiring moxifloxacin treatment. Symptoms disappeared completely. Posterior ToC is negative. Subsequently, sequencing both samples referred to the NRL showed A2059G transition. Conclusion: The clinical cases presented notified the importance of early and accurate diagnosis of MG infections and use of adequate treatment schemes. We emphasized the relevance of monitoring and surveillance prevalence of macrolide-resistant MG in Argentina.

scholarly journals Comparative Study of PCR-Based Approaches for the Genetic Characterization of Three Strains of Acidithiobacillus caldus Isolated from Different Sites in China

2013 ◽  
Vol 62 (4) ◽  
pp. 351-358
Author(s):  
Xueling Wu ◽  
Hong Duan ◽  
Hongwei Fan ◽  
Zhenzhen Zhang ◽  
Lili Liu
Keyword(s):  
16S Rrna ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
23S Rrna ◽  
Rrna Gene ◽  
Sequencing Analysis ◽  
Intergenic Spacers ◽  
Acidithiobacillus Caldus ◽  
Pcr Fingerprinting ◽  
23S Rrna Gene

Comparative study of the genetic characteristics among three Acidithiobacillus caldus strains isolated from different typical environments in China was performed using a combination of molecular methods, namely sequencing analysis of PCR-amplified 16S rRNA genes and 16S-23S rRNA gene intergenic spacers (ITS), repetitive element PCR (rep-PCR), arbitrarily primed PCR (AP-PCR) fingerprinting and random amplified polymorphic DNA (RAPD). Both of the 16S rRNA gene and 16S-23S rRNA gene intergenic spacers sequences of the three strains exhibited small variations, with 99.9-100%, 99.7-100% identity respectively. In contrast, according to the analysis of bacterial diversity based on rep-PCR and AP-PCR fingerprinting, they produced highly discriminatory banding patterns, and the similarity values between them varied from 61.97% to 71.64%. RAPD analysis showed that banding profiles of their genomic DNA exhibited obvious differences from each other with 53.44-75% similarity. These results suggested that in contrast to 16S rRNA genes and 16S-23S rRNA gene intergenic spacers sequencing analysis, rep-PCR, AP-PCR fingerprinting and RAPD analysis possessed higher discriminatory power in identifying these closely related strains. And they could be used as rapid and highly discriminatory typing techniques in studying bacterial diversity, especially in differentiating bacteria within Acidithiobacillus caldus.

Molecular mechanisms of macrolide and fluoroquinolone resistance among Korean isolates of Mycoplasma genitalium over a period of five years 2014–2019

2021 ◽  
Vol 70 (11) ◽  
Author(s):  
Yumi Seo ◽  
Heeyoon Park ◽  
Gilho Lee
Keyword(s):  
Genetic Diversity ◽  
Antimicrobial Resistance ◽  
Type Species ◽  
Mycoplasma Genitalium ◽  
Quinolone Resistance ◽  
23S Rrna ◽  
Rrna Gene ◽  
Content Type ◽  
Link Type ◽  
23S Rrna Gene

Antimicrobial resistance in Mycoplasma genitalium has become a global issue, and certain groups have a higher probability of acquiring resistant strains. Little is known about the genetic diversity and characteristics of the antimicrobial resistance-determining sites (ARDSs) of M. genitalium in the Korean population. Therefore, we examined the genetic diversity of the ARDSs of M. genitalium-positive urogenital samples obtained from Korean females (G1) and males (G2) visiting primary care clinics and DNA samples from referred males (G3) with persistent urethritis. From 2014 to 2019, 54 patients from G1, 86 patients from G2, and 68 patients from G3 were included in the study. Sanger sequencing was performed on the 2058/2059 sites in the 23S rRNA gene and quinolone resistance-determining regions (QRDRs) of M. genitalium . The rates of mutation in G1, G2, and G3 were 1.85, 5.81, and 48.53 %, respectively, for A2059G in the 23S rRNA gene (P<0.001); 1.85, 0, and 17.78 %, respectively, for M95R or I in gyrA (P<0.001); 0, 0, and 31.11 %, respectively, for D99N or G in gyrA (P<0.001); and 7.41, 16.28, and 30 %, respectively, for S83R or N or I in parC (P=0.015). A2059G significantly increased the risk of mutations at the gyrA95, gyrA99, and parC83 sites (all P<0.01). In conclusion, although the genetic diversity of the ARDSs of M. genitalium was variable among the groups, it was generally lower in isolates with macrolide resistance and higher in isolates with quinolone resistance in Korea compared with the isolates in other countries. The G3 group demonstrated increased genetic diversity at the A2059G, gyrA95, gyrA99, and parC83 sites.

scholarly journals Bacterial Load in Daily Urine Samples of Patients Infected withMycoplasma genitalium, Mutation Analysis, and Response to Treatment

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
M. Gossé ◽  
S. A. Nordbø ◽  
B. Pukstad
Keyword(s):  
Mutation Analysis ◽  
Bacterial Load ◽  
False Negative ◽  
Mycoplasma Genitalium ◽  
Urine Samples ◽  
Response To Treatment ◽  
23S Rrna ◽  
Rrna Gene ◽  
Wild Type ◽  
23S Rrna Gene

Objective. Increasing macrolide resistant strains ofMycoplasma genitaliumis a challenge, and to differentiate between treatment failure and reinfection a timely test of cure (TOC) is warranted. The aim of this study was to evaluate the best time for TOC after five days’ treatment ofMycoplasma genitaliuminfection with azithromycin.Methods. Nineteen patients with positive PCR forMycoplasma genitaliumin urine provided urine samples daily for 2 weeks and on days 21, 28, and 35. Samples were tested by a commercial qPCR and by sequencing of the 23S rRNA gene.Results. Eight patients with a wild type ofMycoplasma genitaliumresponded successfully within four days after treatment initiation. Eleven patients had a mutation in the 23S rRNA gene. These samples exhibited high variations in bacterial load, and some patients tested negative at several time points during the observation period.Conclusions. Day-to-day fluctuations in the mutation samples allow for false negative TOC during the first 5 weeks after start of treatment. Due to increasing macrolide resistance ofMycoplasma genitalium, pretreatment mutation analysis is recommended. When a wild type is verified, TOC performed one week after initiation of treatment is suggested.

scholarly journals Clinical case of failure of josamycin in a patient with urethritis caused by Mycoplasma genitalium

2018 ◽  
Vol 94 (4) ◽  
pp. 55-59 ◽  
Author(s):  
L. M. Zubareva ◽  
I. A. Eydel’shteyn ◽  
A. V. Romanov ◽  
V. V. Evstaf’ev ◽  
R. S. Kozlov
Keyword(s):  
Antibiotic Resistance ◽  
Molecular Genetic ◽  
Mycoplasma Genitalium ◽  
23S Rrna ◽  
Routine Practice ◽  
Rrna Gene ◽  
First Line ◽  
Sexually Transmitted ◽  
23S Rrna Gene ◽  
Unknown Status

Mycoplasma genitalium is one of the obligate pathogens that cause sexually transmitted diseases. To detect this pathogen in routine practice, only molecular genetic methods are used that are also used to identify the resistance of MGE to antibiotics. The first-line drugs for the treatment of diseases caused by MGE, are tetracycline and macrolides. In recent years, many countries have increasingly recorded cases of unsuccessful therapy macrolides. Mutations that confer antibiotic resistance to macrolides for Mycoplasm genitalium are concentrated in nucleotide positions 2058 and 2059 in region V of the 23S rRNA gene. Unknown status of macrolide resistance M. genitalium can lead to the development of a persistent infection. We describe the first reported cases of clinical josamycin treatment failure from patient with ure - thritis. The reason for antibiotic resistance was a mutation in the 23S rRNA of MGE as a nucleotide substitution in position A2058G.

Mycoplasma genitalium and its resistance to azithromycin in incarcerated men from Far North Queensland

Sexual Health ◽  
2014 ◽  
Vol 11 (6) ◽  
pp. 587 ◽  
Author(s):  
Gemma Maree Daley ◽  
Darren B. Russell ◽  
Sepehr N. Tabrizi ◽  
Jimmy Twin ◽  
William J. H. McBride
Keyword(s):  
Mycoplasma Genitalium ◽  
Macrolide Resistance ◽  
Urine Samples ◽  
23S Rrna ◽  
Rrna Gene ◽  
Far North ◽  
Incarcerated Men ◽  
23S Rrna Gene

This study examined the prevalence of Mycoplasma genitalium in incarcerated men from Far North Queensland as well as the prevalence of macrolide resistance in identified isolates. Overall, eight out of 140 [5.71% (95% CI 1.82–9.60)] urine samples tested positive and two out of eight (25%) samples carried a mutation in the 23S rRNA gene associated with macrolide resistance.

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

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.

scholarly journals A Novel SimpleProbe PCR Assay for Detection of Mutations in the 23S rRNA Gene Associated with Macrolide Resistance in Mycoplasma genitalium in Clinical Samples

2016 ◽  
Vol 54 (10) ◽  
pp. 2563-2567 ◽  
Author(s):  
Marianne Gossé ◽  
Hilde Lysvand ◽  
Brita Pukstad ◽  
Svein Arne Nordbø
Keyword(s):  
Melting Curve ◽  
Mycoplasma Genitalium ◽  
Curve Analysis ◽  
23S Rrna ◽  
Clinical Samples ◽  
Melting Curve Analysis ◽  
Rrna Gene ◽  
Content Type ◽  
23S Rrna Gene ◽  
Resistant Strains

Macrolide-resistant strains ofMycoplasma genitaliumare an increasing problem throughout the world, and the implementation of a rapid and sensitive assay for mutation detection to guide treatment is needed. Macrolide-resistant strains have been shown to contain base substitutions in positions 2058 and 2059 (Escherichia colinumbering) in region V of the 23S rRNA gene. In this study, we present a SimpleProbe PCR followed by melting curve analysis to differentiate between macrolide-resistant mutants and wild types. The assay was performed on 159Mycoplasma genitalium-positive samples, and the results were compared with DNA sequencing. We also looked at the prevalence of macrolide-resistant strains in a Norwegian population. Of 139 samples characterized successfully by sequencing, 54 (39%) were wild types and 85 (61%) were mutants, consisting of 59 (42%) A2059G, 24 (17%) A2058G, 1 (1%) A2058T, and 1 (1%) A2059C mutation. The melting curve analysis correctly differentiated between wild-type and mutant strains in all cases, but it could not identify the different mutant types. The SimpleProbe PCR proved to be a simple, rapid, and reliable method for the detection of macrolide-resistant isolates ofMycoplasma genitaliumin a clinical setting.

Mycoplasma genitalium acquisition and macrolide resistance after initiation of HIV pre-exposure prophylaxis in men who have sex with men

2020 ◽  
Vol 96 (6) ◽  
pp. 396-398 ◽  
Author(s):  
Jens Tomas Van Praet ◽  
Sanne Steyaert ◽  
Stefaan Vandecasteele ◽  
Barbara Van Den Bergh ◽  
Hilde Mahieu ◽  
...  
Keyword(s):  
Mycoplasma Genitalium ◽  
Macrolide Resistance ◽  
23S Rrna ◽  
Rrna Gene ◽  
Younger Age ◽  
23S Rrna Gene ◽  
Sex With Men ◽  
Taqman Array ◽  
Exposure Prophylaxis

ObjectivesRecent evidence shows that patients using HIV pre-exposure prophylaxis (PrEP) have an increased rate of bacterial STIs, including syphilis, chlamydia and gonorrhoea. Our study aimed to describe the acquisition and the susceptibility for macrolides of Mycoplasma genitalium in men who have sex with men (MSM) on PrEP.MethodsWe studied all MSM who started PrEP in the AZ Sint-Jan Hospital Bruges from 1 June 2017 to 31 March 2019 with at least one follow-up visit. Patients were screened for M. genitalium and other STIs with pooled rectal swabs, pharyngeal swabs and first-voided urine, and blood samples 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 region V of the 23S rRNA gene (A2058G, A2059G, A2058C and others). Patients with an STI were treated based on a national guideline.Results131 MSM (median age 40 years, range 20–79) were included in the study. The median follow-up time was 12 months (IQR 6.1–17). Baseline prevalence of M. genitalium was 6.9% and incidence rate after PrEP initiation was 28.8 per 100 person-years (95% CI 21.7 to 37.2), without significant differences in proportions between the first four quarterly intervals. All but one acquisitions were asymptomatic. Younger age and positivity for M. genitalium at baseline were significantly associated with incident M. genitalium acquisition. The observed proportion of macrolide resistance increased not significantly from 44% at baseline to 57%–86% after PrEP initiation. None of the 27 macrolide-resistant M. genitalium acquisitions could be linked to azithromycin exposure in the three preceding months.ConclusionsAfter initiation of PrEP, we found a stable incidence of almost exclusively asymptomatic M. genitalium. However, a non-significant trend of an increased percentage of macrolide-resistant strains was observed.

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

2016 ◽  
Vol 54 (6) ◽  
pp. 1593-1597 ◽  
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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