Abstract：The rpsl, rrs and embB gene fragments of M. tuberculosis isolates collected throughout Guiyang were PCR-amplified and screened for mutations by direct sequencing methodology. These strains included 19 resistant strains resistant to both streptomycin and ethambutol, and 20 strains susceptible to both streptomycin and ethambutol. Results showed that 73.7% (14/19) resistant M. tuberculosis isolates had mutations in the rpsl gene, and the rpsl AAG43AGG substitution predominated (85.7%). One isolate had mutations in the rpsl AAG43AAT, one in the rpsl AAG88AGG. The rpsl AAG43AGG substitution was also found in one susceptible strain, and mutation rate was 5.0% in 20 susceptible strains. No mutations in the rrs gene were found in both resistant and susceptible strains. Of 84.2%(16/19) resistant isolates had mutations in the embB gene, including embB ATG306GTG substitution in 6 strains, ATG306ATT, ATG306ATC and CAG497CGG in 3 strains respectively. Three strains harboured the embB GGC406GCC substitution and 3 had GGC406AGC mutation. No mutation of embB fragment was determined in all the 20 susceptible strains. The results showed diversity in the rpsl and embB mutations related with streptomycin or ethambutol in M. tuberculosis in Guiyang. The rpsl43 and embB306 are the predominant mutation loci.
孙荣, 欧维正, 王燕, 蒙俊, 秦万, 毕雅坤, 陈峥宏. 贵阳市结核分枝杆菌链霉素和乙胺丁醇耐药相关基因突变的检测与分析[J]. 中国人兽共患病学报, 2016, 32(8): 760-764.
SUN Rong, OU Wei-zheng, WANG Yan, MENG Jun, QING Wan, BI Ya-kun, CHEN Zheng-hong. Detection of mutations associated with streptomycin and ethambutol resistance in Mycobacterium tuberculosis clinical isolates. Chinese Journal of Zoonoses, 2016, 32(8): 760-764.
[1] World Health Organization(WHO): Global Tuberculosis Report[R], Geneva:WHO press.2014. [2] Wang LX, Cheng SM, Chen MT, et al. The fifth national tuberculosis epidemiological survey in 2010[J]. Chin J Antitube rculosis, 2012, 34(8): 485-508. (in Chinese) 王黎霞, 成诗明, 陈明亭, 等. 2010年全国第五次结核病流行病学抽样调查报告[J]. 中国防痨杂志, 2012, 34(8):485-508. [3] Cuevas-Córdoba B, Cuellar-Sánchez A, Pasissi-Crivelli A, et al. rrs and rpsl mutations in streptomycin-resistant isolates of Mycobacterium tuberculosis from Mexico[J]. J Microbiol Immunol Infect. 2013, 46(1): 30-34. doi:10.1016/j.jmii.2012.08.020 [4] Sirgel FA, Warren RM, Streicher EM, et al. EmbB 306 mutations as molecular indicators to predict ethambutol susceptibilit y in Mycobacterium tuberculosis [J]. Chemotherapy, 2012, 58(5): 358-363.doi:10.1159/000343474 [5] Canetti G, Fox W, Khomenko A, et al. Advances in techniques of testing mycobacterial drug sensitivity, and the use of se nsitivity tests in tuberculosis control programmes[J]. Bull World Health Organ, 1969, 41(1): 21-43. [6] Wang Y, Xie JP, Hu CH,et al. Molecular epidemiological research and rapid detection of streptomycin-resistant gene in drug-resist ant Mycobacterium tuberculosis [J]. Chin J Infect, 2006, 6(5): 313-316. (in Chinese) 王莹, 谢建平, 胡昌华等. 耐药结核分枝杆菌链霉素耐药基因的分子流行病学研究及其快速检测 [J]. 中国感染与化疗杂志, 2006, 6 (5): 313-316. [7] Loots du T. An altered Mycobacterium tuberculosis metabolome induced by katG mutations resulting in isoniazid resistan ce[J]. Antimicrob Agents Chemother, 2014, 58(4): 2144-2149. doi:10.1128/AAC.02344-13 [8] Tseng ST, Tai CH, Li CR, et al. The mutations of katG and inhA genes of isoniazid-resistant Mycobacterium tuberculosis is olates in Taiwan[J]. J Microbiol Immunol Infect, 2015, 48(3): 249-255. doi:10.1016/j.jmii.2013.08.018 [9] Dalla Costa ER, Ribeiro MO, Silva MS, et al. Correlations of mutations in katG , oxyR-ahpC and inhA genes and in vitro susceptibility in Mycobacterium tuberculosis clinical strains segregated by spoligotype families from tuberculosis prevalent cou ntries in South America[J]. BMC Microbiol, 2009, 9: 39. doi:10.1186/1471-2180-9-39 [10] Xu G, Wang J, Guo Y, et al. Analysis of the molecular evolution of the Mycobacterium tuberculosis drug-resistant gene rpoB in Asia using a Bayesian evolutionary method[J]. Clin Lab, 2015, 61(8): 1017-1025. [11] Bi YK, Ou WZ, Luo KW, et al. Isoniazid resistance and related gene mutation of 31 Mycobacterium tuberculosis clinical isolates in Guiyang area[J]. Chin J Pathogen, 2014, 9(10): 885-887.DOI: (in Chinese) 毕雅坤, 欧维正, 骆科文,等. 31株结核分枝杆菌异烟肼耐药相关基因检测及序列分析[J]. 中国病原生物学杂志, 2014, 9(10): 885-887. [12] Ou WZ, Chen ZH, Chen J, et al. Determination of resistance genes katG and inhA in Mycobacterium tuberculosis isola tes from Guizhou Province by gene chip[J]. Chin J Zoonoses, 2014, 9(10): 885-887. DOI:(in Chinese) 欧维正, 陈峥宏, 陈静, 等. 基因芯片技术检测贵州省结核分枝杆菌耐药基因 katG 和 inhA [J]. 中国人兽共患病学报, 2015, 31 (7): 655-658. [13] Ou WZ, Luo KW, Chen ZH, et al. Analysis of characteristic of ropB gene mutations associated with Mycobacterium tu bercu losis resistance to rifampicin in Guizhou of China[J]. Chin J Clin Pharmacol, 2015, 31(10): 833-835.DOI: (in Chinese) 欧维正, 骆科文, 陈峥宏, 等. 贵州地区结核分枝杆菌利福平耐药相关基因ropB突变特征分析[J]. 中国临床药理学杂志, 2015, 31(10): 833-835. [14] Shi R, Zhang J, Li C, et al. Detection of streptomycin resistance in Mycobacterium tuberculosis clinical isolates from C hina as determined by denaturing HPLC analysis and DNA sequencing[J]. Mic Infect, 2007, 9(14/15): 153 8-1544. [15] Sun YJ, Luo JT, Wong SY, et al. Analysis of rpsl and rrs mutations in Beijing and non-Beijing streptomycin-resistant Mycobacterium tuberculosis isolates from Singapore[J]. Clin Mic Infect, 2010, 16(3): 287-289.doi:10.1111/j.1469-0691 [16] Potdar P, Thakur P. Development of sequence based molecular diagnostic test to evaluate MDR and XDR in M. tuberculosis patients from Western India[J]. Am J Infect Dis Microbiol, 2013, 1(3): 50-58. [17] Meier A, Sander P, Schaper KJ, et al. Correlation of molecular resistance mechanisms and phenotypic resistance levels in streptomycin-resistant Mycobacterium tuberculosis [J]. Antimicrob Agents Chemother, 1996, 40(11): 2452-2454. [18] Ramaswamy S, Amin AG, Goksel S, et al. Molecular genetic analysis of nucleotide polymorphisms associated with etha mbutol resistance in human isolates of Mycobacterium tuberculosis [J]. Antimicrob Anents Chemother, 2000, 44(2): 321. [19] Zhang H, Chen X, Wang Z, et al. Pyrosequencing analysis for mutations in embB codon306 among clinical Mycobacterium tuberculosis isolates from Qingdao, China[J]. Int J Clin Exp Med, 2015, 8(7): 11276-11282 [20] Zhao LL, Sun Q, Liu HC, et al. Analysis of embCAB mutations associated with ethambutol resistance in multidrug-resist ant Mycobacterium tuberculosis isolates from China[J]. Antimicrob Agents Chemother, 2015, 59(4): 2045-2050.doi:10.1128/ AAC.04933-14
2016, Vol. 32 
