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韩国脓肿分枝杆菌临床菌株中克拉霉素诱导耐药菌株的检测与评估:PCR方法

Detection and assessment of clarithromycin inducible resistant strains among Korean Mycobacterium abscessus clinical strains: PCR methods.

作者信息

Lee Seung-Heon, Yoo Hee Kyung, Kim Seol Hee, Koh Won-Jung, Kim Chang Ki, Park Young Kil, Kim Hee Jin

机构信息

Korean Institute of Tuberculosis, Chungbuk, Korea.

出版信息

J Clin Lab Anal. 2014 Sep;28(5):409-14. doi: 10.1002/jcla.21702. Epub 2014 Mar 20.

Abstract

BACKGROUND

Mycobacterium abscessus group belongs to a group of rapidly growing mycobacteria (RGM) and, following Mycobacterium avium complex, is the second most common pathogen responsible for lung disease caused by nontuberculous mycobacteria (NTM). Clarithromycin is known to be the key drug in the treatment of M. abscessus group disease, but a high failure rate of treatment response is reported due to clarithromycin inducible resistance.

METHODS

Using the results from a clarithromycin susceptibility test we examined the proportion of clarithromycin inducible resistant M. abscessus (sensu stricto; hereafter referred to as M. abscessus) clinical strains. Also, we attempted to detect the clarithromycin resistant strains, using the amplification refractory mutation system-PCR (ARMS-PCR) and real-time PCR methods for rapid detection of single-nucleotide polymorphisms (SNPs) at position 28 (T or C) of the erm(41) gene of M. abscessus leading to resistance to clarithromycin.

RESULTS

Of the 157 M. abscessus clinical strains, clarithromycin susceptible, resistant, and inducible resistant strains accounted for 10.83% (n = 17), 22.29% (n = 35), and 66.88% (n = 105), respectively. Clarithromycin resistant strains were able to separate from clarithromycin susceptible strains by ARMS-PCR and real-time PCR identical to DNA sequence analysis.

CONCLUSION

Most M. abscessus clinical strains in Korea are resistant to clarithromycin, and ARMS-PCR and real-time PCR are useful tools for the rapid detection of single-nucleotide polymorphisms (SNPs) at position 28 of the erm(41) gene.

摘要

背景

脓肿分枝杆菌组属于快速生长分枝杆菌(RGM),是继鸟分枝杆菌复合群之后,引起非结核分枝杆菌(NTM)肺病的第二常见病原体。已知克拉霉素是治疗脓肿分枝杆菌组疾病的关键药物,但据报道,由于克拉霉素诱导耐药,治疗反应的失败率很高。

方法

利用克拉霉素敏感性试验结果,我们检测了克拉霉素诱导耐药的脓肿分枝杆菌(狭义;以下简称脓肿分枝杆菌)临床菌株的比例。此外,我们尝试使用扩增阻滞突变系统PCR(ARMS-PCR)和实时PCR方法检测克拉霉素耐药菌株,以快速检测脓肿分枝杆菌erm(41)基因第28位(T或C)的单核苷酸多态性(SNP),该SNP导致对克拉霉素耐药。

结果

在157株脓肿分枝杆菌临床菌株中,克拉霉素敏感、耐药和诱导耐药菌株分别占10.83%(n = 17)、22.29%(n = 35)和66.88%(n = 105)。通过ARMS-PCR和实时PCR能够将克拉霉素耐药菌株与克拉霉素敏感菌株区分开,结果与DNA序列分析相同。

结论

韩国大多数脓肿分枝杆菌临床菌株对克拉霉素耐药,ARMS-PCR和实时PCR是快速检测erm(41)基因第28位单核苷酸多态性(SNP)的有用工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bfe6/6807423/a08703d83450/JCLA-28-409-g001.jpg

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