title: Molecular characterization of fluoroquinolone-resistant Mycobacterium tuberculosis clinical isolates from Shanghai, China.
abstract: China is one of the countries with the highest prevalence of fluoroquinolone-resistant (FQ(r)) Mycobacterium tuberculosis. Nevertheless, knowledge on the molecular characterization of the FQ(r)M. tuberculosis strains of this region remains very limited. This study was performed to investigate the frequencies and types of mutations present in FQ(r)M. tuberculosis clinical isolates collected in Shanghai, China. A total of 206 FQ(r)M. tuberculosis strains and 21 ofloxacin-sensitive (FQ(s)) M. tuberculosis strains were isolated from patients with pulmonary tuberculosis in Shanghai. The phenotypic drug susceptibilities were determined by the proportion method, and the mutations inside quinolone resistance-determining region (QRDR) of gyrA and gyrB genes were identified by DNA sequence analyses. Among 206 FQ(r)M. tuberculosis strains, 44% (90/206) were multidrug-resistant isolates and 39% (81/206) were extensively drug-resistant isolates. Only 9% (19/206) were monoresistant to ofloxacin. In total, 79.1% (163/206) of FQ(r) isolates harboured mutations in either gyrA or gyrB QRDR. Mutations in gyrA QRDR were found in 75.7% (156/206) of FQ(r) clinical isolates. Among those gyrA mutants, a majority (75.6%) harboured mutations at amino acid position 94, with D94G being the most frequent amino acid substitution. Mutations in gyrA QRDR showed 100% positive predictive value for FQ(r)M. tuberculosis in China. Mutations in gyrB were observed in 15.5% (32/206) of FQ(r) clinical isolates. Ten novel mutations were identified in gyrB. However, most of them also harboured mutations in gyrA, limiting their contribution to FQ(r) resistance in M. tuberculosis. Our findings indicated that, similar to other geographic regions, mutations in gyrA were shown to be the major mechanism of FQ(r) resistance in M. tuberculosis isolates. The mutations in gyrA QRDR can be a good molecular surrogate marker for detecting FQ(r)M. tuberculosis in China.
authors.affiliation: Shanghai Public Health Clinical Center Affiliated to Fudan University, Shanghai, China.
authors.forename: Cuiyun
authors.initials: C
authors.lastname: Zhu
authors.forename: Yongxin
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yinzhong
authors.initials: Y
authors.lastname: Shen
authors.forename: Gilman Kit Hang
authors.initials: GK
authors.lastname: Siu
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Wu
authors.forename: Xueqin
authors.initials: X
authors.lastname: Qian
authors.forename: Guilin
authors.initials: G
authors.lastname: Deng
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
authors.forename: Ricky
authors.initials: R
authors.lastname: Lau
authors.forename: Xiaoyong
authors.initials: X
authors.lastname: Fan
authors.forename: Wenhong
authors.initials: W
authors.lastname: Zhang
authors.forename: Hongzhou
authors.initials: H
authors.lastname: Lu
authors.forename: Wing-Cheong
authors.initials: WC
authors.lastname: Yam
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D002681:China; D027081:DNA Gyrase / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D024841:Fluoroquinolones / Q000494:pharmacology*; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D020125:Mutation, Missense; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D017354:Point Mutation; D017422:Sequence Analysis, DNA; D018088:Tuberculosis, Multidrug-Resistant / Q000382:microbiology; D014397:Tuberculosis, Pulmonary / Q000382:microbiology*
chemical_list: D000995:Antitubercular Agents; D024841:Fluoroquinolones; D027081:DNA Gyrase