Rapid detection of mutations in erm(41) and rrl associated with clarithromycin resistance in Mycobacterium abscessus complex by denaturing gradient gel electrophoresis

被引:8
作者
Liu, Weijia [1 ]
Li, Bing [2 ]
Chu, Haiqing [2 ,5 ]
Zhang, Zhemin [2 ]
Luo, Liulin [3 ]
Ma, Wei [4 ]
Yang, Shiyi [1 ]
Guo, Qi [1 ]
机构
[1] Tongji Univ, Sch Med, Shanghai 200092, Peoples R China
[2] Tongji Univ, Sch Med, Shanghai Pulm Hosp, Dept Resp Med, 507 Zhengmin Rd, Shanghai 200433, Peoples R China
[3] Tongji Univ, Sch Med, Shanghai Pulm Hosp, Dept Clin Lab Med, Shanghai 200433, Peoples R China
[4] Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China
[5] Tongji Univ, Sch Med, Shanghai Pulm Hosp, Shanghai Key Lab TB, Shanghai 200433, Peoples R China
基金
中国国家自然科学基金;
关键词
Mycobacterium abscessus; Clarithromycin susceptibility; erm(41); rrl; Denaturing gradient gel electrophoresis; NONTUBERCULOUS MYCOBACTERIA; TREATMENT OUTCOMES; SUBSP ABSCESSUS; RPOB MUTATIONS; MASSILIENSE; GENE; TUBERCULOSIS; IDENTIFICATION; DIAGNOSIS; BOLLETII;
D O I
10.1016/j.mimet.2017.10.010
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Clarithromycin resistance is increasing dramatically among Mycobacterium abscessus complex. The main resistance mechanisms are mutations in the erm(41) and rrl genes. Here we report PCR-based denaturing gradient gel electrophoresis (DGGE) as an alternative method for rapidly detection of mutations in erm(41) and rrl among M. abscessus isolates. Four primer sets targeting the full-length erm(41) gene and a 354 bp fragment of the rrl gene were designed. A combination of 16 different DGGE patterns were observed for erm(41) gene, including 16 in M. abscessus subsp. abscessus and 1 in M. abscessus subsp. massiliense. Six DGGE patterns were obtained for rrl gene. Mutations in the erm(41) and rrl detected by DGGE were 100% identical to mutations detected by DNA sequencing. This is the first report to identify PCR-based DGGE as a practical, relatively inexpensive technique for rapidly detecting mutations in the erm(41) and rrl genes associated with clarithromycin resistance in M. abscessus complex.
引用
收藏
页码:87 / 93
页数:7
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