2022－2023年内蒙古自治区羊种布鲁氏菌多位点序列分型与体外抗生素敏感性试验

左柯铭; 塔娜; 高静; 关娜; 刘晓婷; 李鹏; 白雪晶; 温永俊

doi:10.3784/jbjc.202312130660

2022－2023年内蒙古自治区羊种布鲁氏菌多位点序列分型与体外抗生素敏感性试验

Multilocus sequence typing and in vitro antibiotic susceptibility testing of Brucella melitensis isolated in the Inner Mongolia Autonomous Region, 2022−2023

摘要

摘要:
目的通过生化鉴定与分子生物型分型鉴定临床分离得到的布鲁氏菌，并测定羊种布鲁氏菌分离株对常见抗生素的体外敏感性。
方法对29株2022—2023年内蒙古自治区（内蒙古）收集到的羊种布鲁氏菌进行生化鉴定、牛种羊种猪种犬种（AMOS）多重荧光定量聚合酶链式反应（PCR）、多位点序列分型（MLST），并选择了阿奇霉素、多西环素、利福平、环丙沙星、左氧氟沙星、链霉素、庆大霉素、头孢曲松共8种抗生素使用E-test药敏试纸条测定各菌株对抗生素的最小抑菌浓度（MIC）。
结果 AMOS多重荧光定量PCR与生化鉴定结果得到羊种1型9株、羊种2型6株、羊种3型14株，ST分型结果为MLST 8型28株、ST 12型1株。全部菌株对多西环素、链霉素、环丙沙星、左氧氟沙星、庆大霉素均敏感；20.68%的菌株对头孢曲松耐药；6.89%的菌株对利福平耐药；100.00%的菌株对阿奇霉素耐药，且MIC范围较其他地区显著增加，这种耐药性可能与地理分布或时间因素有关。
结论利福平作为治疗布病的一线药物，耐药菌株的出现提示，应加强对利福平耐药性的监测工作，链霉素、环丙沙星、左氧氟沙星、庆大霉素均可能作为利福平耐药布病患者治疗的候选药物。

Abstract:
Objective To identify the clinical isolates of Brucella by biochemical identification and molecular bio-typing, and test in vitro susceptibility of Brucella melitensis (B. melitensis) isolates to common antibiotics by E-test drug susceptibility test.
Methods Biochemical identification, abortus melitensis ovis suis(AMOS), multiplex polymerase chain reaction (PCR) and multilocus sequence typing (MLST) were performed for 29 B. melitensis strains collected in the Inner Mongolia Autonomous Region form 2022 to 2023. Eight antibiotics, including azithromycin, doxycycline, rifampicin, ciprofloxacin, levofloxacin, streptomycin, gentamicin and ceftriaxone, were selected to test their minimum inhibitory concentration (MIC) to the strains using E-test strips. By comparing with the MIC reports of B. melitensis collected in different areas, the possible change in drug resistance of strains was determined.
Results AMOS multiple fluorescence quantitative PCR and biochemical identification detected 9 strains of B.melitensis biovar 1, 6 strains of B.melitensis biovar 2 and 14 strains of B.melitensis biovar 3. The MLST typing results showed 28 strains of MLST 8 and 1 strain of MLST 12. All the strains were sensitive to doxycycline, streptomycin, ciprofloxacin, levofloxacin and gentamicin, 20.68% of the strains were resistant to ceftriaxone, 6.89% strains were resistant to rifampicin, and 100.00% of the strains were resistant to azithromycin, and the MIC range increased significantly compared with other areas. This resistance might be related to geographic distribution or time factors.
Conclusion Rifampicin is the first choice for the treatment of brucellosis. The emergence of drug-resistant strains reminds us to strengthen the monitoring of rifampicin resistance. Streptomycin, ciprofloxacin, levofloxacin and gentamicin might be candidates for the treatment of rifampicin-resistant brucellosis patients.

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