应用4种方法检测结核分枝杆菌临床分离株对链霉素的耐受性.pdfVIP

遗 传 学 报 Acta Genetica Sinica , July 2006, 33 (7)：655–663 ISSN 0379-4172 Detection of Streptomycin Resistance in Mycobacterium tu- berculosis Clinical Isolates Using Four Molecular Methods in China 1,① 1 1 1 2 WU Xue-Qiong , LU Yang , ZHANG Jun-Xian , LIANG Jian-Qin , ZHANG Guang-Yu , LI 1 2 3 Hong-Min , LÜ Cui-Huan , DING Bei-Chuan 1. Tuberculosis Research Laboratory, Tuberculosis Center, the Second Affiliated Hospital, Chinese PLA General Hospital, Beijing 100091, China; 2. Thorax Disease Hospital of Hebei Province, Shijiazhuang 050041, China; 3. The Research Institute of TB Control and Prevention of Beijing,Beijing 100035, China Abstract: To evaluate the relationship between mutations in rpsL or rrs genes and streptomycin (SM) resistance, we compared four molecular methods for their clinical value in the detection of SM resistance. Genotypic analysis of SM resistance in 167 M. tuber- culosis clinical strains isolated from Chinese patients was performed by direct DNA sequencing, SSCP, RFLP, and reverse dot-blot hybridization (RDBH) assays. Of the 98 SM-resistant isolates, 78 (79.6%) had missense mutations in codon 43 or 88 of rpsL re- sulting in a Lys to Arg substitution, 6 (6.1%) had mutations of the rrs gene at positions 513 A to C or T or 516 C to T, and 14 (14.3%) had the wild-type sequence. None of the 69 SM-susceptible isolates examined had alterations in rpsL or rrs. The results of the SSCP, RFLP, and RDBH analyses for these mutations and wild-type sequences were completely consistent with DNA sequenc- ing data. Five distinct single-nucleotide substitutions in codon 43 or 88 of rpsL gene or in