Mutations in ParC and GyrA of moxifloxacin-resistant and susceptible Mycoplasma genitalium strains

Macrolide or fluoroquinolone-resistant Mycoplasma genitalium is spreading worldwide. We aimed to determine the influence of single nucleotide polymorphisms (SNPs) in the quinolone resistance determining regions (QRDR) of parC and gyrA in cultured M. genitalium strains. In addition, we examined the prevalence of macrolide- and fluoroquinolone resistance mediating mutations in specimens collected from Japanese male patients with urethritis in two time-periods between 2005–2009 and 2010–2017, respectively, by sequencing the QRDR of parC and gyrA and domain V of the 23S rRNA gene. The minimum inhibitory concentrations (MIC) of moxifloxacin, sitafloxacin, ciprofloxacin, levofloxacin, doxycycline, minocycline, azithromycin and clarithromycin were determined in 23 M. genitalium strains. Three cultured strains had elevated MICs for moxifloxacin at 16, 4 and 2 mg/L and had SNPs with the amino-acid change Ser83→Ile in ParC (p<0.001) and 3 kinds of SNPs with amino-acid changes Asp99→Asn, Gly93→Cys and Met95→Ile in GyrA, respectively. Among a total of 148 M. genitalium positive urine specimens, the prevalence of A2058G and A2059G SNPs in the 23S rRNA gene and any SNPs in ParC increased from 4.8% and 22.6% in 2005–2009 to 42.2% and 53.1% in 2010–2017, respectively. If M. genitalium is considered multi-drug resistant in clinical specimens carrying SNPs in the 23S rRNA gene and Ser83→Ile in ParC, the prevalence of multi-drug resistance is 12.5% in 2010–2017 in Japan. In conclusion, the SNP resulting in Ser83→Ile in ParC is closely related to moxifloxacin resistance even though other factors may also affect treatment outcomes by moxifloxacin. The prevalence of circulating multi-drug resistant M. genitalium strains with macrolide- and fluoroquinolone-resistance is dramatically increasing in Japan.

[1]  L. G. Góes,et al.  Macrolide Resistance – associated 23 S rRNA Mutation in Mycoplasma genitalium , Japan , 2019 .

[2]  M. Unemo,et al.  In Vitro Activity of Sitafloxacin and Additional Newer Generation Fluoroquinolones Against Ciprofloxacin-Resistant Neisseria gonorrhoeae Isolates. , 2018, Microbial drug resistance.

[3]  R. Hamasuna,et al.  The detection of microorganisms related to urethritis from the oral cavity of male patients with urethritis. , 2017, Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy.

[4]  S. Nakagawa,et al.  Proposed Pharmacokinetic-Pharmacodynamic Breakpoint of Garenoxacin and Other Quinolones. , 2017, Japanese journal of infectious diseases.

[5]  M. Yasuda,et al.  Emergence of Mycoplasma genitalium with clinically significant fluoroquinolone resistance conferred by amino acid changes both in GyrA and ParC in Japan. , 2017, Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy.

[6]  S. Garland,et al.  Increasing Macrolide and Fluoroquinolone Resistance in Mycoplasma genitalium , 2017, Emerging infectious diseases.

[7]  M. Cusini,et al.  Background review for the 2016 European guideline on Mycoplasma genitalium infections , 2016, Journal of the European Academy of Dermatology and Venereology : JEADV.

[8]  T. Hachisuga,et al.  The first report: An analysis of bacterial flora of the first voided urine specimens of patients with male urethritis using the 16S ribosomal RNA gene-based clone library method. , 2016, Microbial pathogenesis.

[9]  T. Ezaki,et al.  Remarkable increase in fluoroquinolone-resistant Mycoplasma genitalium in Japan. , 2014, The Journal of antimicrobial chemotherapy.

[10]  J. Jensen,et al.  Mycoplasma genitalium testing pattern and macrolide resistance: a Danish nationwide retrospective survey. , 2014, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[11]  M. Unemo,et al.  In Vitro Activity of the New Fluoroketolide Solithromycin (CEM-101) against Macrolide-Resistant and -Susceptible Mycoplasma genitalium Strains , 2014, Antimicrobial Agents and Chemotherapy.

[12]  G. Gilbert,et al.  Failure of moxifloxacin treatment in Mycoplasma genitalium infections due to macrolide and fluoroquinolone resistance , 2013, International journal of STD & AIDS.

[13]  M. Yasuda,et al.  P2.100 Clinical Efficacy of Sitafloxacin 100Mg Twice Daily For 7 Days For Patients with Non-Gonococcal Urethritis , 2013, Sexually Transmitted Infections.

[14]  R. Hamasuna Mycoplasma genitalium in male urethritis: Diagnosis and treatment in Japan , 2013, International journal of urology : official journal of the Japanese Urological Association.

[15]  G. Gilbert,et al.  Fluoroquinolone and Macrolide Resistance-Associated Mutations in Mycoplasma genitalium , 2013, Journal of Clinical Microbiology.

[16]  M. Yasuda,et al.  Clinical efficacy of sitafloxacin 100 mg twice daily for 7 days for patients with non-gonococcal urethritis , 2013, Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy.

[17]  R. Hamasuna Identification of treatment strategies for Mycoplasma genitalium-related urethritis in male patients by culturing and antimicrobial susceptibility testing , 2013, Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy.

[18]  D. Taylor-Robinson,et al.  Mycoplasma genitalium: from Chrysalis to Multicolored Butterfly , 2011, Clinical Microbiology Reviews.

[19]  M. Yasuda,et al.  Macrolide Resistance–associated 23S rRNA Mutation in Mycoplasma genitalium, Japan , 2011, Emerging Infectious Diseases.

[20]  C. Mcgowin,et al.  Mycoplasma genitalium: An Emerging Cause of Sexually Transmitted Disease in Women , 2011, PLoS pathogens.

[21]  M. Yasuda,et al.  Effect of gatifloxacin against Mycoplasma genitalium-related urethritis: an open clinical trial , 2011, Sexually Transmitted Infections.

[22]  L. Lopez,et al.  Re-evaluating the treatment of nongonococcal urethritis: emphasizing emerging pathogens--a randomized clinical trial. , 2011, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[23]  T. Tsukamoto,et al.  Clinical efficacy of levofloxacin 500 mg once daily for 7 days for patients with non-gonococcal urethritis , 2011, Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy.

[24]  M. Yasuda,et al.  Macrolide resistance-associated 23S rRNA mutation in Mycoplasma genitalium, Japan. , 2011, Emerging infectious diseases.

[25]  M. Yasuda,et al.  Emergence of clinical strains of Mycoplasma genitalium harbouring alterations in ParC associated with fluoroquinolone resistance. , 2010, International journal of antimicrobial agents.

[26]  J. Jensen,et al.  Antimicrobial Susceptibilities of Mycoplasma genitalium Strains Examined by Broth Dilution and Quantitative PCR , 2009, Antimicrobial Agents and Chemotherapy.

[27]  T. Mroczkowski,et al.  A randomized comparison of azithromycin and doxycycline for the treatment of Mycoplasma genitalium-positive urethritis in men. , 2009, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[28]  S. Tabrizi,et al.  Azithromycin treatment failure in Mycoplasma genitalium-positive patients with nongonococcal urethritis is associated with induced macrolide resistance. , 2008, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[29]  J. Jensen,et al.  Isolation of Mycoplasma genitalium from First-Void Urine Specimens by Coculture with Vero Cells , 2007, Journal of Clinical Microbiology.

[30]  S. Garland,et al.  Azithromycin Failure in Mycoplasma genitalium Urethritis , 2006, Emerging infectious diseases.

[31]  J. Jensen,et al.  Antibiotic Susceptibility Testing of Mycoplasma genitalium by TaqMan 5′ Nuclease Real-Time PCR , 2005, Antimicrobial Agents and Chemotherapy.

[32]  M. Morozumi,et al.  Emergence of Macrolide-Resistant Mycoplasma pneumoniae with a 23S rRNA Gene Mutation , 2005, Antimicrobial Agents and Chemotherapy.

[33]  J. Jensen,et al.  Use of TaqMan 5′ Nuclease Real-Time PCR for Quantitative Detection of Mycoplasma genitalium DNA in Males with and without Urethritis Who Were Attendees at a Sexually Transmitted Disease Clinic , 2004, Journal of Clinical Microbiology.

[34]  J. Jensen,et al.  Tetracycline treatment does not eradicate Mycoplasma genitalium , 2003, Sexually transmitted infections.

[35]  M. Yasuda,et al.  Association of Mycoplasma genitalium Persistence in the Urethra With Recurrence of Nongonococcal Urethritis , 2001, Sexually transmitted diseases.

[36]  P. Hannan,et al.  Guidelines and recommendations for antimicrobial minimum inhibitory concentration (MIC) testing against veterinary mycoplasma species. International Research Programme on Comparative Mycoplasmology. , 2000, Veterinary research.

[37]  J. Jensen,et al.  Isolation of Mycoplasma genitalium strains from the male urethra , 1996, Journal of clinical microbiology.