Evaluation of a rapid detection method of clarithromycin resistance genes in Mycobacterium avium complex isolates
Evaluation of a rapid detection method of clarithromycin resistance genes in Mycobacterium avium complex isolates
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DOI：
10.1093/jac/dkq536
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发表时间：
2011-04-01
影响因子：
5.2
通讯作者：
Ogawa, Kenji
中科院分区：
文献类型：
--
作者：
Inagaki, Takayuki;Yagi, Tetsuya;Ogawa, Kenji
目的：克拉霉素是禽分枝杆菌复合体（MAC）疾病各种治疗方案中的关键药物，也是唯一一种药物敏感性与这些疾病临床反应相关的药物。据报道，在高水平克拉霉素耐药分离株中，23S rRNA基因2058或2059位点发生点突变。在这项研究中，我们检验了药敏试验结果与MAC耐药相关的23S rRNA基因突变之间的相关性。此外，我们采用了扩增难解突变系统(ARMS)-PCR快速检测方法来鉴定MAC分离株的23S rRNA基因突变。方法：采用基于NCCLS/CLSI推荐的微量稀释法，测定245株临床MAC分离株克拉霉素的MIC。对其中219株克拉霉素敏感株和26株克拉霉素耐药株进行23S rRNA基因测序和ARMS-PCR分析。结果：药敏试验显示耐药株和敏感株mic均呈双峰分布。23S rRNA基因序列分析显示，所有克拉霉素敏感菌株均为野生型，24株克拉霉素耐药菌株为突变型。序列和ARMS-PCR分析的敏感性分别为92.3%和84.6%，特异性均为100%。结论：我们发现克拉霉素mic与23S rRNA基因突变之间存在相关性。应用ARMS-PCR检测MAC分离株的23S rRNA突变，可用于克拉霉素耐药性的快速检测。
Objectives: Clarithromycin is the key drug in the various treatment regimens of Mycobacterium avium complex (MAC) diseases, and is the only drug for which drug susceptibility has been shown to correlate with clinical response in these diseases. A point mutation at either positions 2058 or 2059 of the 23S rRNA gene has been reported to occur in high-level clarithromycin-resistant isolates. In this study, we examined the correlation between the results from a drug susceptibility test and the mutation of the 23S rRNA gene involved in drug resistance in MAC. Furthermore, we adapted a rapid detection method using amplification refractory mutation system (ARMS)-PCR to identify mutations in the 23S rRNA gene in MAC isolates.Methods: Using a microdilution method based on the NCCLS/CLSI recommendation, the MIC of clarithromycin was determined for 245 clinical MAC isolates. Of these, 219 clarithromycin-susceptible and 26 clarithromycin-resistant strains were analysed by sequencing of the 23S rRNA gene and ARMS-PCR.Results: The drug susceptibility test revealed a bimodal distribution of MICs for both the susceptible and resistant strains. Sequence analysis of the 23S rRNA gene revealed that all of the clarithromycin-susceptible strains were wild-type whereas 24 of the clarithromycin-resistant strains were mutant type. The sensitivity of the sequence and ARMS-PCR analyses was 92.3% and 84.6%, respectively, and the specificity of both was 100%.Conclusions: We found a correlation between MICs of clarithromycin and 23S rRNA gene mutations. ARMS-PCR for 23S rRNA mutations of MAC isolates is useful for rapid detection of clarithromycin resistance.