Çalışkan Zeynep Cansu, Tanrıverdi Elif Seren, Uzun Mertcan, Otlu Barış, Zarakolu Pınar
Hacettepe University Faculty of Medicine, Department of Infectious Diseases and Clinical Microbiology, Ankara, Türkiye.
İnönü University Faculty of Medicine, Department of Medical Microbiology, Malatya, Türkiye.
Mikrobiyol Bul. 2024 Oct;58(4):353-365. doi: 10.5578/mb.20249624.
A sexually transmitted bacterium, Mycoplasma genitalium has varying rates of reported resistance to macrolide and some fluoroquinolone group antimicrobials recommended for the treatment of its infections. It is currently recommended that the treatment of these must be planned according to macrolide resistance status. The aim of this study was to determine the presence of macrolide resistance associated mutations (MRM) and fluoroquinolone resistance associated mutations (QRM) in patients infected with M.genitalium. Sixty-one patients who were ≥ 18 years old, presented to our outpatient clinic between March 2017-March 2022, had symptoms of urethritis/cervicitis according to the Centers for Disease Control and Prevention definition were included in the study. By nucleic acid amplification test (NAAT), the presence of M.genitalium (Mycoplasma-Ureaplasma-OSR for BD MAX, BioGX, the Netherlands) as well as Neisseria gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis (BDMAX system, BD Diagnostics, USA) in the first stream urine samples was determined. Patients' age, gender, sexual orientation if indicated, diagnostic test results for human immunodeficiency virus (HIV) and syphilis, history of antibiotic use in the last three months, presence of concomitant microorganisms detected by NAAT and urine culture results of the symptomatic period were also recorded. Urine samples in which M.genitalium was detected were stored at -80 °C until the study day. On the study day, they were thawed and a modified real-time polymerase chain reaction (Rt-PCR) test was performed targeting the V region (147 bp) of the 23S rRNA gene for MRM and gyrA (nucleotides 172-402), gyrB (nucleotides 1256-1480), parC (nucleotides 164-483) and parE (nucleotides 1210-1489) gene regions for QRM. IBM SPSS 25 (IBM Inc., Armonk, NY, USA) software was used for descriptive statistical analysis of the patient data. Of the patients; 49 were male, 12 were female. The age range was 20-57 years. Sexual orientation of 15 (30.6%) male patients was men who have sex with men (MSM). Sixteen (26.2%) were individuals living with HIV and 14 (87.5%) were MSM. Four patients had previous syphilis infection. By NAAT, a second microorganism was present in 30 patients with M.genitalium; Ureaplasma urealyticum in 27 (90%), C.trachomatis in two (6.7%) and N.gonorrhoeae in one (3.3%) patient. Urine cultures performed in 42 (68.8%) of 61 patients during the symptomatic period yielded Lactobacillus delbrueckii in one patient. Eighteen (29.5%) patients had a history of antimicrobial use in the last three months. Macrolide resistance associated mutations was detected in 45 (73.8%) and QRM in 20 (32.8%) of M.genitalium infected individuals. Of those with MRM, 17 (37.8%) had concurrent QRM. Macrolide resistance associated mutations were detected at positions A2071G (75.6%) and A2072G (24.4%) in the 23S rRNA gene. The presence of QRM was detected in parC (85%) and gyrA regions (15%). C234T mutation in parC was detected in nine patients (45%), C184T in four, A248T in three and A248A in one, while A288G mutation in gyrA was detected in two patients and G285T mutation in one. Chi-square test showed no significant correlation between the presence of mutations associated with resistance and MSM, HIV/syphilis infection status and antimicrobial use in the last three months (p> 0.05). To the best of our knowledge, our study is the first study on antimicrobial resistance of M.genitalium in Türkiye and emphasizes the importance of macrolide resistance in symptomatic patients infected with M.genitalium. Further studies are needed for the clinical significance of mutations associated with fluoroquinolone resistance. Determination of antimicrobial resistance in M.genitalium diagnostic tests will be useful in terms of guiding treatment and preventing inappropriate treatment approaches in the early period.
生殖支原体是一种性传播细菌,对推荐用于治疗其感染的大环内酯类和一些氟喹诺酮类抗菌药物的耐药率各不相同。目前建议,必须根据大环内酯类耐药情况来规划对这些感染的治疗。本研究的目的是确定感染生殖支原体的患者中与大环内酯类耐药相关的突变(MRM)和与氟喹诺酮类耐药相关的突变(QRM)的存在情况。纳入研究的61例患者年龄≥18岁,于2017年3月至2022年3月期间到我们的门诊就诊,根据疾病控制与预防中心的定义有尿道炎/宫颈炎症状。通过核酸扩增试验(NAAT),确定首段尿液样本中生殖支原体(用于BD MAX、BioGX、荷兰的支原体-脲原体-OSR)以及淋病奈瑟菌、沙眼衣原体和阴道毛滴虫(BDMAX系统,BD诊断公司,美国)的存在情况。还记录了患者的年龄、性别、性取向(如表明)、人类免疫缺陷病毒(HIV)和梅毒的诊断检测结果、过去三个月的抗生素使用史、NAAT检测到的伴随微生物的存在情况以及症状期的尿液培养结果。检测到生殖支原体的尿液样本在-80°C下保存至研究日。在研究日,将样本解冻,并针对MRM的23S rRNA基因的V区域(147 bp)以及QRM的gyrA(核苷酸172 - 402)、gyrB(核苷酸1256 - 1480)、parC(核苷酸164 - 483)和parE(核苷酸1210 - 1489)基因区域进行改良实时聚合酶链反应(Rt-PCR)检测。使用IBM SPSS 25(IBM公司,美国纽约州阿蒙克)软件对患者数据进行描述性统计分析。患者中,49例为男性,12例为女性。年龄范围为20 - 57岁。15名(30.6%)男性患者的性取向为男男性行为者(MSM)。16名(26.2%)为HIV感染者,其中14名(87.5%)为MSM。4例患者曾有梅毒感染。通过NAAT,在30例感染生殖支原体的患者中存在第二种微生物;27例(90%)为解脲脲原体,2例(6.7%)为沙眼衣原体,1例(3.3%)为淋病奈瑟菌。61例患者中有42例(68.8%)在症状期进行了尿液培养,其中1例培养出德氏乳杆菌。18例(29.5%)患者在过去三个月有抗菌药物使用史。在感染生殖支原体的个体中,45例(73.8%)检测到与大环内酯类耐药相关的突变,20例(32.8%)检测到QRM。在有MRM的患者中,17例(37.8%)同时存在QRM。在23S rRNA基因的A2071G(75.6%)和A2072G(24.4%)位置检测到与大环内酯类耐药相关的突变。在parC(85%)和gyrA区域(15%)检测到QRM的存在。在parC中检测到9例患者(45%)有C234T突变,4例有C184T突变,3例有A248T突变,1例有A248A突变,而在gyrA中检测到2例患者有A288G突变,1例有G285T突变。卡方检验显示,耐药相关突变的存在与MSM、HIV/梅毒感染状态以及过去三个月的抗菌药物使用之间无显著相关性(p>0.05)。据我们所知,我们的研究是土耳其关于生殖支原体抗菌药物耐药性的首次研究,强调了大环内酯类耐药在感染生殖支原体的有症状患者中的重要性。对于与氟喹诺酮类耐药相关的突变的临床意义,还需要进一步研究。在生殖支原体诊断检测中确定抗菌药物耐药性将有助于指导治疗并在早期预防不适当的治疗方法。