脓肿分枝杆菌16S rRNA检测及生物学特性研究  

Detection of 16S rRNA gene and biological characteristics of Mycobacterium abscessus

作　　者：孙铭艳[1] 吴倩倩 王业鑫 王楠[1] 刘言霞 陶元勇[1] Sun Mingyan;Wu Qianqian;Wang Yexin;Wang Nan;Liu Yanxia;Tao Yuanyong(Clinical Laboratory,Affiliated Hospital of Weifang Medical University,Shandong 261031,China;Department of Joint Surgery,Affiliated Hospital of Weifang Medical University,Shandong 261031,China)

机构地区：[1]潍坊医学院附属医院检验科,山东261031 [2]潍坊医学院附属医院关节外科,山东261031

出　　处：《中华皮肤科杂志》2018年第12期901-904,共4页Chinese Journal of Dermatology

摘　　要：目的鉴定1株脓肿分枝杆菌临床分离株。方法取2017年12月18日就诊于潍坊医学院附属医院的1例疑似非结核分枝杆菌感染患者的脓液标本行细菌培养、革兰染色和抗酸染色;药敏试验采用比例法。提取菌株基因组DNA,选用16S rRNA通用引物进行PCR扩增,回收、纯化产物后测序,与GenBank数据库中已知脓肿分枝杆菌序列比较同源性。采用基质辅助激光解吸电离飞行时间质谱鉴定分离株。结果基质辅助激光解吸电离飞行时间质谱与16S rRNA基因检测均鉴定该分离菌为脓肿分枝杆菌。药敏试验显示该菌对阿米卡星、莫西沙星、左氧氟沙星敏感,对链霉素、异烟肼、利福平、乙胺丁醇、氧氟沙星、卡那霉素、卷曲霉素、对氨基水杨酸、丙硫异烟胺、利福布汀耐药。诊断为左膝关节皮下软组织感染。根据药敏试验结果,给予阿米卡星和左氧氟沙星治疗后,症状好转。结论16S rRNA检测与基质辅助激光解吸电离飞行时间质谱可用于脓肿分枝杆菌鉴定。Objective To identify a clinical isolate of Mycobacterium abscessus.Methods A pus sample was collected from a patient with suspected nontuberculous mycobacterial infection who visited the Affiliated Hospital of Weifang Medical University on December 18,2017,and was subjected to bacterial culture,Gram staining and acid-fast staining.Drug sensitivity test was conducted by the proportion method. The genome DNA of the strain was extracted and amplified by PCR with the universal primer of 16S rRNA. The PCR products were sequenced after collection and purification,and were compared with the known sequence of Mycobacterium abscessus in GenBank database.The isolate was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).Results The clinical isolate was identified as Mycobacterium abscessus both by MALDI-TOF MS and 16S rRNA gene sequencing. The drug sensitivity test showed that the strain was sensitive to amikacin,moxifloxacin,levofloxacin,but was resistant to streptomycin,isoniazid,rifampicin,ethambutol,ofloxacin,kanamycin,capreomycin, aminosalicylic acid,protionamide and rifabutin.The patient was diagnosed with subcutaneous soft tissue infection in the left knee joint.According to the results of drug sensitivity test,the patient was treated with amikacin and levofloxacin,and her condition was improved after treatment.Conclusion The 16S rRNA gene detection and MALDI-TOF MS both can be applied in the identification of Mycobacterium abscessus.

关 键 词：非结核分枝杆菌 RNA 核糖体 16S 光谱法 质量 基质辅助激光解吸电离 微生物敏感性试验 脓肿分枝杆菌 

分 类 号：R446.5[医药卫生—诊断学]