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Phenotypic Antimicrobial Susceptibility and Genotypic Characterization of Clinical Ureaplasma Isolates Circulating in
Hongxia Ma1, Xuemei Zhang2, Xiaoxing Shi3
1Department of Health Management Medicine, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
Abstract:
There is a growing global concern regarding the rise of antimicrobial resistance among Ureaplasma spp. isolates. However, studies on the antimicrobial susceptibility profiles, resistance mechanisms, and clonality of Ureaplasma spp. clinical isolates are still limited and cover only some geographic regions. Firstly, Ureaplasma species from the urogenital tracts of patients in Shanghai, China, were isolated by using the culture medium (A8 and 10B broth), and identified the genotype by polymerase chain reaction (PCR). Secondly, the antimicrobial susceptibility tests were determined by using broth microdilution assay. Then, the resistance genetic determinants to fluoroquinolones (FQs), macrolides, and tetracyclines were investigated through PCR/DNA sequencing. Finally, the molecular epidemiology of Ureaplasma species was studied by multilocus sequence typing (MLST). Among 258 isolates, Ureaplasma parvum (UPA) and Ureaplasma urealyticum (UUR) were found in 226 (87.60%) and 32 (12.40%) isolates, respectively. The minimum inhibitory concentrations (MICs) of 258 Ureaplasma spp. strains ranged from 0.015 to 64μg/ml for all 11 kinds of antimicrobials. Regardless of species, the isolates were most sensitive to AZI (1.94%), JOS (3.49%), and CLA (4.23%). Among them, there were 39 (15.12%) multidrug-resistant (MDR) strains, including 32 UPA isolates. The resistance rates of UPA to CIP (91.59%), and ROX (36.28%) were significantly higher than those of UUR. Twenty six FQ-resistant isolates had amino acid substitutions in gyrA and in parC (Ser83Leu). Mutations were detected in genes encoding ribosomal proteins L4 (Thr84Ile) and L22 (Ser81Pro) in macrolide-resistant isolates. Tet(M) was found in four UPA isolates. These mutations were mainly found in UPA isolates. Sequence type 1 (ST1) was the predominant ST, which contained 18 isolates. In conclusion, this study showed a higher resistance rate (especially to ROX and CIP), higher substitution rate, and higher MDR rate among UPA strains. The most active antimicrobial agents were AZI, JOS, and CLA. Identifying UPA or UUR in clinical isolates could help clinicians to choose appropriate drugs for treatment. The main resistance mechanisms may involve gene substitution of Ser83Leu in parC and Ser81Pro in L22. ST1 was the predominant ST of Ureaplasma isolates with MDR to FQs and macrolides in Shanghai, China.
Insights
Antimicrobial resistance in Ureaplasma species is a growing concern, particularly in Ureaplasma parvum (UPA) which shows higher resistance rates to certain antibiotics like ciprofloxacin and roxithromycin. Identifying Ureaplasma parvum or Ureaplasma urealyticum is crucial for effective treatment selection.
Area of Science:
- Microbiology
- Infectious Diseases
- Molecular Epidemiology
Background:
- Rising global antimicrobial resistance (AMR) in Ureaplasma spp. necessitates further investigation.
- Limited data exists on AMR profiles, resistance mechanisms, and clonality of Ureaplasma clinical isolates across different regions.
Purpose of the Study:
- To investigate the antimicrobial susceptibility, resistance mechanisms, and molecular epidemiology of Ureaplasma spp. clinical isolates from Shanghai, China.
- To compare resistance patterns between Ureaplasma parvum (UPA) and Ureaplasma urealyticum (UUR).
Main Methods:
- Isolation and identification of Ureaplasma species using culture media and PCR.
- Antimicrobial susceptibility testing via broth microdilution.
- Investigation of genetic determinants for resistance to fluoroquinolones, macrolides, and tetracyclines using PCR/DNA sequencing.
- Multilocus sequence typing (MLST) for molecular epidemiology.
Main Results:
- Ureaplasma parvum (UPA) was identified in 87.60% and Ureaplasma urealyticum (UUR) in 12.40% of 258 isolates.
- Azithromycin (AZI), josamycin (JOS), and clarithromycin (CLA) showed the highest sensitivity.
- Multidrug-resistant (MDR) strains accounted for 15.12%, predominantly UPA.
- UPA exhibited significantly higher resistance rates to ciprofloxacin (CIP) and roxithromycin (ROX) compared to UUR.
- Specific mutations in gyrA, parC, ribosomal proteins L4 and L22, and the tet(M) gene were identified as resistance mechanisms, mainly in UPA.
- Sequence type 1 (ST1) was the predominant clone among MDR isolates.
Conclusions:
- Ureaplasma parvum strains demonstrate higher resistance rates, substitution rates, and MDR rates, particularly to ROX and CIP.
- Azithromycin, josamycin, and clarithromycin are the most effective antimicrobial agents against these isolates.
- Identifying UPA or UUR is critical for guiding appropriate clinical treatment decisions.
- Key resistance mechanisms involve specific gene substitutions in parC and ribosomal protein L22.
- ST1 represents the dominant Ureaplasma clone associated with MDR to fluoroquinolones and macrolides in Shanghai.
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