脓肿分枝杆菌临床分离株抗生素敏感性与基因型的关系

Relationship between antibiotic susceptibility and genotype of Mycobacterium abscessus clinical isolates

目的观察脓肿分枝杆菌临床分离株抗生素敏感性与基因型的关系。方法选择2016年1月~2018年9月上海市肺科医院分离出的脓肿分枝杆菌分离株87株进行研究,其中痰标本72株,支气管肺泡灌洗液15株。所有菌株均经MGIT960培养基生长和对硝基苯甲酸试验鉴定为非结核分枝杆菌,通过对rPOB基因测序进一步鉴定为脓肿分枝杆菌。对所有菌株进行定量聚合酶链式反应扩增,采用NCBI Nucleotide blast program进行单核苷酸多态性分析,采用微量稀释法测定抗生素敏感性,分析抗生素敏感性与基因型的关系。结果 87株分离株包括81个subsp.abscessus,6个subsp.massiliense。81个subsp.abscessus中共72个ERM(41)T28序列和9个ERM(41)C28序列,ERM(41)序列对10种抗生素敏感;81个subsp.abscessu亚型中包括41个克拉霉素(CLA)敏感株和10个CLA中介株,8个ERM(41)C28和43个ERM(41)T28分离株不存在rrl 2058/2059突变,30个耐药分离株中包括29个ERM(41)T28分离株(其中5个发生rrl 2058/2059突变)和1个ERM(41)C28分离株(1个发生rrl2058/2059突变)。subsp.massiliense耐药分离株2个,1个发生rrl 2058/2059突变。结论 rrl和ERM(41)基因型对脓肿分离杆菌临床分离株CLA耐药具有一定预测作用,阿米卡星治疗脓肿分枝杆菌可能具有更佳效果,值得进一步研究探讨。

Objective To observe the relationship between antibiotic susceptibility and genotype of clinical isolates of Mycobacterium abscessus. Methods From January 2016 to September 2018, 87 strains of Mycobacterium abscess isolated from Shanghai Pulmonary Hospital were selected for the study. Among them, 72 strains were sputum and 15 strains were bronchoalveolar lavage fluid. All strains were identified as Nontuberculous mycobacteria by MGIT960 medium growth and p-nitrobenzoic acid test, while further identified as Mycobacterium abscessus by sequencing rPOB gene. All strains were subjected to quantitative polymerase chain reaction amplification, while single nucleotide polymorphism analysis was performed using NCBI Nucleotide blast program. Antibiotic sensitivity was determined by microdilution method, while the relationship between antibiotic sensitivity and genotype was analyzed. Results Eightyseven isolates included 81 subsp.abscessus, 6 subsp.massiliense. A total of 72 ERM(41) T28 sequences and 9 ERM(41) C28 sequences in 81 subsp.abscessus, and ERM(41) sequences to 10 species were sensitivity to antibiotics. 81 subsp.abscessu subtypes included 41 Claroycin(CLA)-sensitive strains and 10 CLA-mediated strains, 8 ERM(41)C28 and 43 ERM(41) T28 isolates without rrl 2058/2059 mutation. 30 resistant isolates were consisted of 29 ERM(41)T28 isolates(5 of them had rrl 2058/2059 mutations) and 1 ERM(41) C28 isolate(1 occurred the rrl 2058/2059 mutation). There were 2 subsp.massiliense resistant isolates, and 1 had a rrl 2058/2059 mutation. Conclusion The rrl and ERM(41) genotypes have a certain predictive effect on CLA resistance of clinical isolates of abscess abacillus. The treatment of Mycobacterium abscessus with Amikacin may have better effect and it is worthy of further study.