临床分离快速生长分枝杆菌的鉴定及对抗结核药物的敏感性分析

Identification of clinical isolates of rapidly growing mycobacteria and analysis of sensitivity to anti-tuberculosis drugs

目的比较不同方法鉴定快速生长分枝杆菌（RGM）的准确性,检测RGM对抗结核药物的敏感性。方法 18株RGM使用DNA微阵列芯片法和基因测序法进行菌株鉴定,用比例法检测对利福平、异烟肼、链霉素、乙胺丁醇、丙硫异烟胺、对氨基水杨酸、卷曲霉素、氧氟沙星和卡那霉素9种抗结核药物的药物敏感性。结果 DNA微阵列芯片法与基因测序结果对偶然分枝杆菌和脓肿分枝杆菌鉴定符合率为100%,对其他菌株的鉴定结果偏差较大。RGM对临床上常用的9种抗结核药物普遍耐药,耐药程度依次为马赛分枝杆菌（9种药物全耐药）、脓肿分枝杆菌（8-9种药物耐药）、新金色分枝杆菌（8种药物耐药）、偶发分枝杆菌（7种药物耐药）、血液分枝杆菌（7种药物耐药）、康赛医院分枝杆菌（6种药物耐药）和富西亚分枝杆菌（4种药物耐药）。结论分子诊断技术在临床实验室的应用意义重大。使用抗结核药物治疗RGM感染患者的风险极大,应该按照RGM药敏折点标准进行药敏试验,以指导临床合理用药。

Objective To compare the accuracy of different methods for identification of rapidly growing mycobacteria（ RGM）,and to detect the sensitivity of RGM isolates to anti-tuberculosis drugs. Methods 18 strains of RGM were identified by DNA microarray and gene sequencing method. The sensitivity for the RGM isolates of 9 anti-tuberculosis drugs,i. e.,rifampicin,isoniazid,streptomycin,ethambutol,protionamide,aminosalicylic acid,capreomycin,ofloxacin and kanamycin,were tested by proportion method. Results The coincidence rate for the identification of DNA microarray and gene sequencing to Mycobacterium fortuitum and Mycobacterium abscessus was 100%,but obvious deviation of identification results for the other species was found. RGM isolates were commonly resistant to the tested 9 anti-tuberculosis drugs. The resistant degree was as below： Mycobacterium massiliense（ all resistant to 9 drugs）,Mycobacterium abscessus（ resistant to 8-9 drugs）,Mycobacterium neoaurum（ resistant to 8 antibiotics）,Mycobacterium fortuitum（ resistant to 7 drugs）,Mycobacterium septicum（ resistant to 7 drugs）,Mycobacterium conceptionense（ resistant to 6 grugs） and Mycobacterium phocaicum（ resistant to 4 drugs）. Conclusion Molecular diagnostic technique should be of value in clinical laboratory. The treatment of anti-tuberculosis drugs for RGM infection may be greatly risky. The drugs breakpoints of rapid growing Mycobacterium should be adopted to guide the clinical application of anti-tuberculosis drugs.