结核分枝杆菌喹诺酮类耐药基因的分析-免疫学专业论文.docxVIP

PAGE PAGE 4 广州医学院硕士学位论文 Methods A total of 67 MTB clinical strains were originally isolated from patients with pulmonary tuberculosis. The phenotype of susceptibility of each strain was determined by the absolute concentration method and the sequences of the QRDR in gyrA and gyrB genes were detected with DNA direct sequencing technique. Results 47 of 64(73.4%) levofloxacin-resistant MTB clinical isolates, including 20 of 22(90.9%) high-level resistant strains and 27 of 42(64.3%) low-level resistant strains, had mutation in QRDR of gyrA gene. Among the 47 mutated isolates, 45 mutated only in one codon and 2 mutated in double codons. The mutation in single codon was found in codon 70, 89, 90, 91 or 94 and the mutation in double codons was found in codons 90 and 94. Frequency of the mutation in codon 94 was higher than that in other codons and amounted to 63.8%. The mutation patterns in our study involved seven patterns of single codon mutation (H70R, D89N, A90V, S91A, D94G, D94A or D94N) and one pattern of double codons mutation (A90V with D94A). Among the seven patterns of single codon mutation in our data, two ones (H70R and D89N) only developed high-level resistance to levofloxacin. In contrast with the single codon mutation, the double codons mutation is relatively rare and frequently correlated to high-level resistance to levofloxacin. Among 64 levofloxacin-resistant MTB clinical isolates, only one strain mutated in codon 511 of gyrB gene and resulted in a change of ACC→AAC (Thr→Asn). Conclusions Our results confirm that mutations in gyrA and gyrB genes are the major mechanism of quinolone resistance in MTB and suggest that different mutation can lead to different level of quinolone resistance. Nevertheless, mutations in the gyrA and gyrB genes could not account for all quinolone-resistant MTB isolates. Some other mechanisms such as the presence of drug efflux pumps need to be further studied. KEY WORDS: gyrA gene gyrB gene Mycobacterium tuberculosis quinolone