Fluoroquinolone Resistance-associated Mutations Research Articles

Estimation of antimicrobial resistance of Mycoplasma genitalium, Belgium, 2022.

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.

High Genetic Diversity in Third-Generation Cephalosporin-Resistant Escherichia coli in Wastewater Systems of Schleswig-Holstein.

The spread of multidrug-resistant bacteria from humans or livestock is a critical issue. However, the epidemiology of resistant pathogens across wastewater pathways is poorly understood. Therefore, we performed a detailed comparison of third-generation cephalosporin-resistant Escherichia coli (3GCREC) from wastewater treatment plants (WWTPs) to analyze dissemination pathways. A total of 172 3GCREC isolated from four WWTPs were characterized via whole genome sequencing. Clonal relatedness was determined using multi-locus sequence typing (MLST) and core genome MLST. Resistance genotypes and plasmid replicons were determined. A total of 68 MLST sequence types were observed with 28 closely related clusters. Resistance genes to eight antibiotic classes were detected. In fluoroquinolone-resistant isolates, resistance was associated with three-or-more point mutations in target genes. Typing revealed high genetic diversity with only a few clonal lineages present in all WWTPs. The distribution paths of individual lines could only be traced in exceptional cases with a lack of enrichment of certain lineages. Varying resistance genes and plasmids, as well as fluoroquinolone resistance-associated point mutations in individual isolates, further corroborated the high diversity of 3GCREC in WWTPs. In total, we observed high diversity of 3GCREC inside the tested WWTPs with proof of resistant strains being released into the environment even after treatment processes.

563. Mycoplasma genitalium with Fluoroquinolone Resistance: Design and Development of a Multiplex Real-Time PCR Assay for Detection and Differentiation

Abstract Background Mycoplasma genitalium is a sexually transmitted bacterium causing urethritis in men and associated with cervicitis and pelvic inflammatory disease in women. Only a few antimicrobial classes have activity against mycoplasmas. The first-line treatment for M. genitalium infections is doxycycline followed by azithromycin. Due to overuse of azithromycin to treat infections, resistance to azithromycin has been rapidly increasing. The moxifloxacin of fluoroquinolones family of antibiotics was 100% effective against M. genitalium, but recently the resistance markers for fluoroquinolone treatment have also emerged. Multiple studies attribute fluoroquinolone treatment failure to single nucleotide polymorphisms (SNPs) in parC and gyrA genes. We aim to demonstrate proof of principle for a Real-Time PCR assay that simultaneously detects M. genitalium and fluoroquinolone resistance. Methods The MGB Alert M. genitalium with fluoroquinolone resistance RUO Detection Reagent is a multiplex of real-time PCR reagents that simultaneously detect M. genitalium DNA and distinguish fluoroquinolone resistance-associated mutations and wild type in M. genitalium. The assay targets parC and gyrA genes and includes a set of two primers and two probes specific to M. genitalium. The DSQ probe identifies the species DNA. The Pleiades hybridization probes serve to identify and distinguish a wild type genotype and several known resistance-conferring point mutations by melt curve analysis. Results Analytical Study: Analytical sensitivity was determined and verified at 1.42 copy of bacterial genome per reaction. A linear range was established from 1e7 to 10 genome copies per reaction. Test results showed no cross-reactivity with organisms that might be present in normal vaginal swabs and urine specimens. Conclusion Preliminary analytical evaluation of the MGB Alert M. genitalium with fluoroquinolone resistance RUO Detection Reagent indicates potential as a valuable tool in the diagnosis of M. genitalium and resistance guided treatment. Disclosures Boris Alabyev, PhD, ELITechGroup MDx LLC: Salary Irina Ankoudinova, MS, ELITechGroup MDx LLC: Salary Noah Scarr, BS, ELITechGroup MDx LLC: Salary Becca Decker, BS, ELITechGroup MDx LLC: Salary Eugene Lukhtanov, PhD, ELITechGroup MDx LLC: Salary

Changes in prevalence of Mycoplasma genitalium macrolide and fluoroquinolone resistance: notes from a 13-year retrospective survey in Dalarna County, Sweden

ObjectiveMacrolide 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...

Hi-plex deep amplicon sequencing for identification, high-resolution genotyping and multidrug resistance prediction of Mycobacterium leprae directly from patient biopsies by using Deeplex Myc-Lep.

Expansion of antimicrobial resistance monitoring and epidemiological surveillance are key components of the WHO strategy towards zero leprosy. The inability to grow Mycobacterium leprae invitro precludes routine phenotypic drug susceptibility testing, and only limited molecular tests are available. We evaluated a culture-free targeted deep sequencing assay, for mycobacterial identification, genotyping based on 18 canonical SNPs and 11 core variable-number tandem-repeat (VNTR) markers, and detection of rifampicin, dapsone and fluoroquinolone resistance-associated mutations in rpoB/ctpC/ctpI, folP1, gyrA/gyrB, respectively, and hypermutation-associated mutations in nth. The limit of detection (LOD) was determined using DNA of M.leprae reference strains and from 246 skin biopsies and 74 slit skin smears of leprosy patients, with genome copies quantified by RLEP qPCR. Sequencing results were evaluated versus whole genome sequencing (WGS) data of 14 strains, and versus VNTR-fragment length analysis (FLA) results of 89 clinical specimens. The LOD for sequencing success ranged between 80 and 3000 genome copies, depending on the sample type. The LOD for minority variants was 10%. All SNPs detected in targets by WGS were identified except in a clinical sample where WGS revealed two dapsone resistance-conferring mutations instead of one by Deeplex Myc-Lep, due to partial duplication of the sulfamide-binding domain in folP1. SNPs detected uniquely by Deeplex Myc-Lep were missed by WGS due to insufficient coverage. Concordance with VNTR-FLA results was 99.4% (926/932 alleles). Deeplex Myc-Lep may help improve the diagnosis and surveillance of leprosy. Gene domain duplication is an original putative drug resistance-related genetic adaptation in M.leprae. EDCTP2 programme supported by the European Union (grant number RIA2017NIM-1847 -PEOPLE). EDCTP, R2Stop: Effect:Hope, The Mission To End Leprosy, the Flemish Fonds Wetenschappelijk Onderzoek.

Prevalence of fluoroquinolone resistance-associated mutations in Mycoplasma genitalium among clients of two sexual health centres in the Netherlands: a cross-sectional study

ObjectiveThis study aimed to determine the prevalence of fluoroquinolone resistance-associated mutations (QRAMs) in Mycoplasma genitalium (MG) among clients of two sexual health centres (SHCs) in the Netherlands.DesignA cross-sectional study.Setting and...

Clinical Performance of Three Commercial Molecular Diagnostic Assays for the Detection of Fluoroquinolone Resistance-Associated Mutations in Mycoplasma genitalium.

The high prevalence of macrolide resistance in Mycoplasma genitalium results in an increased reliance on moxifloxacin, the second-line treatment; however, moxifloxacin resistance has also emerged. Because assays that can detect fluoroquinolone resistance-associated mutations will be useful for the management of macrolide-resistant M. genitalium infections, we evaluated the performance of three commercial assays (the Allplex MG & MoxiR Assay [Seegene], LightMix Modular parC kit [TIBMOLBIOL], and MGMO qPCR [NYtor) in comparison with parC gene Sanger sequencing used as the reference. Between January 2018 and December 2020, remnants of M. genitalium-positive clinical specimens received at the French National Reference Center for Bacterial Sexually Transmitted Infections were collected if a Sanger sequencing result was obtained for the parC gene. Overall, 368 M. genitalium-positive specimens were assessed. The clinical sensitivities for the detection of the ParC mutations that are likely of clinical significance were 91.8% (95% CI = 83.2 to 96.2), 98.6% (95% CI = 92.4 to 99.8), and 94.4% (95% CI = 86.6 to 97.8) for the Allplex MG & MoxiR, LightMix Modular parC, and MGMO qPCR kits, respectively, with no significant difference between the three kits. The clinical specificity of the Allplex MG & MoxiR and MGMO qPCR kits was 100% (95% CI = 97.7 to 100 and 98.7 to 100, respectively), which was significantly higher than the specificity of the LightMix Modular parC kit of 95.4% (95%CI = 92.3 to 97.3), for which the interpretation of melting curves may be misleading. These kits should be useful for the selection of antimicrobials in macrolide-resistant M. genitalium infections, although further developments may be necessary because parC mutations involved in fluoroquinolone resistance have not been precisely determined.

Prevalence of macrolide and fluoroquinolone resistance-associated mutations in Mycoplasma genitalium in metropolitan and overseas France

ObjectiveLimited macrolide and fluoroquinolone resistance data are available in France for Mycoplasma genitalium. We performed a multicentre cross-sectional study to investigate the prevalence of macrolide and fluoroquinolone resistance-associated mutations in...

Molecular Typing Reveals Distinct Mycoplasma genitalium Transmission Networks among a Cohort of Men Who Have Sex with Men and a Cohort of Women in France.

Mycoplasma genitalium causes sexually transmitted infecti.ons in men and women. Treatment failures to macrolides and fluoroquinolones have been reported worldwide. Although the mgpB typing method has often been used in M. genitalium-infected men who have sex with men (MSM), limited typing data are available for M. genitalium-infected women. In this study, we aimed to investigate the genetic relationship between M. genitalium strains and their antibiotic resistance profile in a cohort of MSM (86.2% on HIV preexposure prophylaxis [PrEP], 13.8% HIV positive) and a large cohort of women using mgpB/MG309 typing. The mgpB types were determined in 374 samples from 305 women and 65 MSM. Three MSM and one woman had two concurrent or subsequent samples. Macrolide and fluoroquinolone resistance-associated mutations were searched in the 23S rRNA as well as parC and gyrA genes. The mgpB phylogenetic construction revealed three large clusters that differed according to sexual practices and geographical origin of patients. The prevalence of macrolide and fluoroquinolone resistance was significantly higher in MSM compared with women (95.4% vs. 14.1% and 30.6% vs. 7.2%, p < 0.001, respectively). The macrolide resistance spread was polyclonal in both populations, but clonal diffusion of two dual-resistant types was observed in PrEP users in association with high antibiotic pressure and dense connectivity in this population.

Antimicrobial resistance in Neisseria gonorrhoeae and Mycoplasma genitalium isolates from the private healthcare sector in South Africa: A pilot study.

Reports have emerged globally of antimicrobial resistance (AMR) in Neisseria gonorrhoeae and Mycoplasma genitalium infections. In South Africa (SA), there are substantial differences between private and public healthcare with regard to antimicrobial drug prescribing practice, which could affect AMR patterns of private and public healthcare patients. To perform a pilot study to determine the frequency of AMR of N. gonorrhoeae and M. genitalium in patients accessing SA's private healthcare sector. In this cross-sectional study, N. gonorrhoeae-positive cultures and M. genitalium DNA samples were collected from a private healthcare reference laboratory from August 2018 to August 2019. In N. gonorrhoeae-positive cultures, antimicrobial susceptibility testing was performed, followed by N. gonorrhoeae multiantigen sequence typing (NG-MAST) to determine genetic relatedness of the isolates. To determine macrolide and fluoroquinolone resistance rates, M. genitalium-positive samples were analysed by sequencing the 23S rRNA, gyrA and parC genes. Twenty-one N. gonorrhoeae- and 27 M. genitalium-positive specimens were included in this analysis. High rates of resistance were detected among gonococcal isolates, with 90% resistance to tetracycline, 86% to penicillin and 62% to ciprofloxacin, but no resistance to azithromycin, cefixime and ceftriaxone. NG-MAST revealed genetically diverse isolates with 83% novel NG-MAST sequence types. Macrolide and fluoroquinolone resistance-associated mutations were detected in 18.5% (n=5/27) and 7.4% (n=2/27) of M. genitalium strains, respectively. We observed high frequencies of ciprofloxacin, penicillin and tetracycline resistance in N. gonorrhoeae and macrolide resistance-associated mutations in M. genitalium in private healthcare sector patients in SA. This finding highlights the need to use diagnostics for sexually transmitted infections and to include the private healthcare sector in antimicrobial surveillance and stewardship programmes.

An alarming high prevalence of resistance-associated mutations to macrolides and fluoroquinolones in Mycoplasma genitalium in Belgium: results from samples collected between 2015 and 2018

ObjectivesThe number of reported cases of multiresistant Mycoplasma genitalium (MG) is increasing globally. The aim of this study was to estimate the prevalence of macrolide and possible fluoroquinolone resistance-associated mutations...

Prevalence of macrolide and fluoroquinolone resistance-associated mutations in Mycoplasma genitalium in Hong Kong

Background: Mycoplasma genitalium (MG) is a common cause of non-gonococcal urethritis (NGU) and the surge in MG antibiotics resistance is alarming. In Hong Kong, NGU is the most prevalent sexually transmitted infection (STI) in males. However, the local prevalence of MG and the burden of antimicrobial resistance remain unknown. This study aimed to determine the prevalence of antibiotic resistance in MG among STI male patients in Hong Kong. Methods and materials: Male patients attending a sexually transmitted diseases clinic for treatment of NGU and presenting with urethritis were included. First-void urine specimens were collected and screened for MG infection by real-time PCR (Genesig, United Kingdom). M. genitalium-positive samples were tested for macrolide resistance-associated mutations in the 23S rRNA gene using the ResistancePlus MG assay (SpeeDx, Australia), and fluoroquinolone resistance-associated genetic markers in gyrA and parC by PCR and sequencing. Results: The prevalence of MG in male patients with symptomatic urethritis was 10.8% (17/157). Among 17 men with MG infection, 5 (29.4%) were initially treated with azithromycin and 4 (23.5%) on moxifloxacin. Overall, 11 patients (64.7%) harbored macrolide resistance-associated mutations in the 23S rRNA gene. The GyrA and ParC genes were successfully amplified and sequenced in 15 MG-positive cases, of which 60.0% (9/15) possessed fluoroquinolone resistance-associated mutations (S83I or D87Y) in parC. The presence of MG with alterations in both 23S rRNA gene and parC, potentially associated with treatment failure with macrolide and fluoroquinolone regimens, was 46.7% (7/15). Conclusion: The emergence of MG carrying both macrolide and fluoroquinolone resistance-associated mutations is raising concern about incurable MG for which recommended therapies would be ineffective. This study indicates further antimicrobial drug resistance surveillance would be needed to limit the spread of macrolide and fluoroquinolone resistance in MG in Hong Kong.

Macrolide and fluoroquinolone associated mutations in Mycoplasma genitalium in a retrospective study of male and female patients seeking care at a STI Clinic in Guangzhou, China, 2016-2018

BackgroundAntimicrobial resistance in M. genitalium is a growing clinical problem. We investigated the mutations associated with macrolide and fluoroquinolone resistance, two commonly used medical regimens for treatment in China. Our aim is to analyze the prevalence and diversity of mutations among M. genitalium-positive clinical specimens in Guangzhou, south China.MethodsA total of 154 stored M. genitalium positive specimens from men and women attending a STI clinic were tested for macrolide and fluoroquinolone mutations. M. genitalium was detected via TaqMan MGB real-time PCR. Mutations associated with macrolide resistance were detected using primers targeting region V of the 23S rRNA gene. Fluoroquinolone resistant mutations were screened via primers targeting topoisomerase IV (parC) and DNA gyrase (gyrA).Results98.7% (152/154), 95.5% (147/154) and 90.3% (139/154) of M. genitalium positive samples produced sufficient amplicon for detecting resistance mutations in 23S rRNA, gyrA and parC genes, respectively. 66.4% (101/152), 0.7% (1/147) and 77.7% (108/139) samples manifested mutations in 23S rRNA, gyrA and parC genes, respectively. A2072G (59/101, 58.4%) and S83I (79/108, 73.1%) were highly predominating in 23S rRNA and parC genes, respectively. Two samples had amino acid substitutions in gyrA (M95I and A96T, respectively). Two samples had two amino acid substitutions in parC (S83I + D87Y). 48.6% (67/138) of samples harbored both macrolide and fluoroquinolone resistance-associated mutations. The most common combination of mutations was A2072G (23S rRNA) and S83I (parC) (40/67, 59.7%). One sample had three amino acid changes in 23S rRNA, gyrA and parC genes (A2072G + A96T + S83I).ConclusionsThe high antimicrobial resistance rate of M. genitalium in Guangzhou is a very worrying problem and suggests that antimicrobial resistance testing and the development of new antibiotic regimens are crucially needed.

Lower mgpB diversity in macrolide-resistant Mycoplasma genitalium infecting men visiting two sexually transmitted infection clinics in Montpellier, France

Men engaged in high-risk sexual behaviour, such as MSM, are likely to be infected by resistant Mycoplasma genitalium strains. Understanding the transmission dynamics is challenging. We aimed to investigate the molecular epidemiology of M. genitalium in men visiting sexually transmitted infection (STI) clinics. Between June 2017 and February 2018, 95 M. genitalium-positive specimens from 78 men, including 76.9% MSM, visiting two STI clinics in Montpellier, France, were analysed for SNPs in the mgpB adhesin gene and number of tandem repeats in the MG_309 gene. Macrolide and fluoroquinolone resistance were determined. Typing results were compared with antibiotic resistance, sexual behaviour, sampling site, HIV pre-exposure prophylaxis (PrEP) usage and HIV status. Thirty-eight mgpB STs were identified, including 23 new STs, with ST4 being most prevalent. The mgpB/MG_309 typing method identified 52 genetic profiles, resulting in a discriminatory index of 0.979. Macrolide and fluoroquinolone resistance-associated mutations were detected in 58.3% and 10.8% of patients, respectively. The macrolide resistance rate was higher among MSM than among men who have sex with women only (68.4% versus 9.1%; adjusted OR, 1.57; 95% CI, 1.13-2.18; P = 0.007). A lower mgpB diversity of 0.870 was found among macrolide-resistant strains in comparison with 0.978 in macrolide-susceptible strains, with an over-representation of mgpB ST62 and ST153. Although macrolide resistance spread appears polyclonal in M. genitalium, the lower diversity of mgpB types among macrolide-resistant strains may reflect the easier spread of a few specific mgpB types or the occurrence of sexual networks among MSM.

Mycoplasma genitalium Infections With Macrolide and Fluoroquinolone Resistance-Associated Mutations in Heterosexual African American Couples in Alabama

Mycoplasma genitalium (MG) is a sexually transmitted pathogen associated with inflammatory syndromes in men and women. Macrolides and fluoroquinolones are recommended MG treatments. The frequency of MG strains with macrolide resistance-associated mutations (MRMs) and quinolone resistance-associated mutations (qRMs) is increasing worldwide, however these data are sparse in populations in the United States. We investigated the prevalence of MG infections with MRMs and qRMs and MG infection concordance within African American couples in Birmingham, AL. We used a real-time polymerase chain reaction to detect MG and identify MRMs. quinolone resistance-associated mutations were detected using traditional polymerase chain reactions amplifying regions in gyrA, gyrB, parC, and parE. The MG concordance in couples was evaluated by MG positivity and MG genotypes. Oral, anal, urine, and/or vaginal specimens were tested from 116 couples. Twenty-eight (12.1%) participants comprising 22 couples tested MG-positive (11.2% in men and 12.9% in women). Macrolide resistance-associated mutations were detected in 17 (60.7%) MG-positive participants, with gender-specific resistance rates of 69.2% for men and 53.3% for women. quinolone resistance-associated mutations were detected in 3 (11.1%) MG-positive participants, all of whom also had MRMs. By MG positivity status, 27.3% of couples were concordant. If MG strain genotypes are also considered, then concordance was 20.0%. Among heterosexual African Americans with MG infection, about 60% had strains with MRMs and 11% had strains with both MRMs and qRMs, highlighting the potential for MG treatment failure to not only macrolides, but also quinolones. These findings may help to guide clinicians in MG testing and treatment decisions in the United States.

Surveillance of the prevalence of macrolide and/or fluoroquinolone resistance-associated mutations in Mycoplasma genitalium in Japan.

To clarify the status of macrolide and fluoroquinolone resistance of clinical strains of Mycoplasma genitalium in Japan, we amplified portions of the gyrA, parC, and 23S rRNA genes from DNAs in 627 first-voided urine specimens collected from men with M.genitalium-positive urethritis who visited clinics mainly in Sendai, Tokyo, and Osaka, Japan, from 2013 to 2017, by PCR and sequenced. The incidence of single amino acid changes at Met95 or Asp99 in GyrA increased chronologically and was approximately 10% from 2015 onward. The incidence of amino acid changes at Ser83 or Asp87 in ParC was approximately 50% in 2013 but increased to 60-70% from 2014 to 2017. The incidence of mutations at A2071 or A2072 in the 23S rRNA gene increased chronologically and reached over 70% in 2017. The prevalence of M.genitalium harboring alterations in ParC and mutations in the 23S rRNA gene increased and was approximately 50% in 2016 and 2017. The prevalence of M.genitalium with alterations in both GyrA and ParC and mutations in the 23S rRNA gene, which could be associated with treatment failures with the sitafloxacin and azithromycin regimens, were approximately 15% and 10% in 2016 and 2017, respectively. The prevalence of M.genitalium with genetic alterations associated with resistance to fluoroquinolones and/or macrolides is increasing rapidly in Japan. We must prevent the further selection of multi-drug-resistant M.genitalium so that M.genitalium infections will not become untreatable.

Tetracycline and fluoroquinolone resistance in clinical Ureaplasma spp. and Mycoplasma hominis isolates in France between 2010 and 2015

As information on Ureaplasma spp. and Mycoplasma hominis resistance is currently limited, the aim of this study was to investigate the susceptibility of Ureaplasma spp. and M. hominis to tetracyclines and fluoroquinolones in France. The susceptibility of 1014 clinical isolates obtained in Bordeaux University Hospital (Bordeaux, France) between 2010 and 2015 was evaluated using two commercial kits, S.I.R. Mycoplasma (Bio-Rad) from 1 January 2010 to 5 October 2012 and MYCOFAST RevolutioN kit (ELITech Group) from 6 October 2012 to 31 December 2015. The MICs of isolates designated as resistant were determined using the broth microdilution assay. Additionally, the tet(M) gene and fluoroquinolone resistance-associated mutations were identified. Among 831 Ureaplasma spp. isolates, the tetracycline, levofloxacin and moxifloxacin resistance rates were 7.5%, 1.2% and 0.1%, respectively. Among 183 M. hominis isolates, the resistance rates were 14.8%, 2.7% and 1.6% for tetracycline, levofloxacin and moxifloxacin, respectively. Over the 6 year period, no significant change in resistance to tetracycline or fluoroquinolones was observed. The tet(M) gene was found in all tetracycline-resistant isolates. All levofloxacin-resistant isolates harboured a mutation in the parC or parE genes. Isolates that were also resistant to moxifloxacin harboured an additional mutation in the gyrA gene. The MYCOFAST RevolutioN kit significantly overestimated levofloxacin and moxifloxacin resistance in Ureaplasma spp. isolates. Resistance to tetracycline and fluoroquinolones is limited in France in mycoplasmas but compared with a previous report in 1999-2002, a significant increase in tetracycline resistance among Ureaplasma spp. was observed. Ongoing monitoring of the antibiotic susceptibility of these urogenital mycoplasmas remains necessary.

Antimicrobial susceptibility of Mycoplasma genitalium isolates from Pretoria, South Africa in 2012 and 2016

Background: Mycoplasma genitalium is a sexually transmitted pathogen associated with non-gonococcal urethritis and cervicitis. Azithromycin regimens have been considered first-line treatment for M. genitalium infections, with fluoroquinolone regimens effective as second-line treatments. However, the proportion of M. genitalium harbouring macrolide or fluoroquinolone resistance-associated mutations has been increasing worldwide. This study was done to compare the genotypic macrolide and fluoroquinolone resistance of M. genitalium strains obtained from women attending a termination of pregnancy clinic five years apart.Methods: M. genitalium was detected by PCR in vaginal swab samples from 100 and 104 termination of pregnancy attendees at a tertiary hospital in Pretoria, South Africa during 2012 and 2016 respectively. Genes associated with macrolide and fluoroquinolone resistance in the M. genitalium isolates were sequenced and analysed.Results: The prevalence of M. genitalium was 6.0% (6/100) in 2012 and 7.7% (8/104) in 2016. No resistance-associated mutations were seen in the 2012 isolates. Among the 2016 M. genitalium isolates, two (25%) harboured a macrolide-associated resistance mutation and one (12.5%) a fluoroquinolone resistance-associated mutation in the parC gene.Conclusions: There is an increase in macrolide and fluoroquinolone resistance among local M. genitalium strains. This highlights the need for improved surveillance.

Antimicrobial susceptibility ofMycoplasma genitaliumisolates from Pretoria, South Africa in 2012 and 2016

AbstractBackground: Mycoplasma genitalium is a sexually transmitted pathogen associated with non-gonococcal urethritis and cervicitis. Azithromycin regimens have been considered first-line treatment for M. genitalium infections, with fluoroquinolone regimens effective as second-line treatments. However, the proportion of M. genitalium harbouring macrolide or fluoroquinolone resistance-associated mutations has been increasing worldwide. This study was done to compare the genotypic macrolide and fluoroquinolone resistance of M. genitalium strains obtained from women attending a termination of pregnancy clinic five years apart.Methods: M. genitalium was detected by PCR in vaginal swab samples from 100 and 104 termination of pregnancy attendees at a tertiary hospital in Pretoria, South Africa during 2012 and 2016 respectively. Genes associated with macrolide and fluoroquinolone resistance in the M. genitalium isolates were sequenced and analysed.Results: The prevalence of M. genitalium was 6.0% (6/100) in 201...

Prevalence of macrolide and fluoroquinolone resistance-mediating mutations in Mycoplasma genitalium in five cities in Russia and Estonia.

Background and objectiveResistance in the sexually transmitted bacterium Mycoplasma genitalium to all recommended therapeutic antimicrobials have rapidly emerged. However, to date, internationally reported resistance surveillance data for M. genitalium strains circulating in Eastern Europe are entirely lacking. The aim of this study was to estimate the prevalence of macrolide and fluoroquinolone resistance-associated mutations in M. genitalium in four cities in Russia and one in Estonia, 2013–2016.Materials and methodsConsecutive urogenital samples found positive for M. genitalium during diagnostic testing were retrospectively analyzed for resistance-associated mutations in the 23S rRNA and parC genes using pyrosequencing and conventional Sanger sequencing, respectively.ResultsIn total, 867 M. genitalium positive samples from 2013–2016 were analyzed. Macrolide resistance-associated mutations were detected in 4.6% of the samples from Russia (0.7–6.8% in different cities) and in 10% of the samples from Estonia. The mutations A2059G and A2058G were highly predominating in both Russia and Estonia, accounting together for 90.9% of the cases positive for nucleotide substitutions in the 23S rRNA gene. The rates of possible fluoroquinolone resistance-associated mutations were 6.2% in Russia (2.5–7.6% in different cities) and 5% in Estonia. The mutations S83I and S83N were the most frequent ones in Russia (24.4% each), whereas D87N highly predominated in Estonia (83.3% of all fluoroquinolone resistance-associated mutations). Approximately 1% of the samples in both countries harbored both macrolide and possible fluoroquinolone resistance-associated mutations, with A2058G and S83I being the most frequent combination (37.5%).ConclusionsThe prevalence of macrolide and fluoroquinolone resistance-associated mutations in M. genitalium was 4.6% and 6.2%, respectively, in Russia, and 10% and 5%, respectively, in Estonia. Despite the relatively low rates of macrolide and fluoroquinolone resistance in these countries, antimicrobial resistance surveillance and testing for resistance-associated mutations in M. genitalium positive cases would be valuable.