A rapid and non-pathogenic assay for association of Mycobacterium tuberculosis gyrBA mutations and fluoroquinolone resistance using recombinant Mycobacterium smegmatis
A rapid and non-pathogenic assay for association of Mycobacterium tuberculosis gyrBA mutations and fluoroquinolone resistance using recombinant Mycobacterium smegmatis
复制标题
使用重组耻垢分枝杆菌快速、非致病性测定结核分枝杆菌 gyrBA 突变与氟喹诺酮耐药性之间的关系
DOI：
10.1093/femsle/fny266
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发表时间：
2018
影响因子：
2.1
通讯作者：
Hoshino Yoshihiko
中科院分区：
文献类型：
--
作者：
Yoshida Mitsunori;Nakata Noboru;Miyamoto Yuji;Fukano Hanako;Ato Manabu;Hoshino Yoshihiko
我们建立了一种重组卡介苗(BCG)和重组污垢分枝杆菌(Mtb)gyrBA基因突变与氟喹诺酮类药物耐药相关的方法。野生型Mtb gyrBA重组菌对FQ的最小抑菌浓度(MIC)与内生型gyrBA相当。在27个gyrBA突变中，以FQ MIC形式的折叠改变为主。斯马蒂斯和M。BovisBCG的背景是可比的，部分等同于之前报道的重组Mtb株。GyrA基因第90或94位突变可产生强烈和协同的FQ耐药性，这可能与临床观察携带这些突变的菌株最常见或次之有关。在这项研究中测试的FQS中，水合西塔沙星的MIC最低，这与之前的活体动物研究结果相似。在临床分离的结核分枝杆菌中检测到的大多数BBA突变可能会导致FQ耐药，但有几个突变会降低细菌的生长速度。总体而言，重组M。Smegmatis似乎是评估毒力分枝杆菌FQ敏感性的一种有益的替代系统。
We developed a method involving recombinantMycobacterium bovisbacillus Calmette–Guérin (BCG) and recombinantMycobacterium smegmatisto determine which mutations inMycobacterium tuberculosis(Mtb)gyrBAare associated with fluoroquinolone (FQ) resistance. The minimal inhibitory concentration (MIC) for FQ for recombinant strains with wild-typeMtb gyrBAwas equivalent to that for strains with intrinsicgyrBA. Among 27gyrBAmutations, the fold-changes in FQ MIC forM. smegmatisandM. bovisBCG backgrounds were comparable and were in part equivalent to those previously reported for recombinantMtbstrains. Mutations at position 90 or 94 ofgyrAconferred strong and synergistic FQ resistance, which may be associated with the clinical observation that isolates carrying these mutations are the most or second most frequent. Sitafloxacin hydrate had the lowest MIC among the FQs tested in this study, which is similar to findings from a previousin vivoanimal study. MostgyrBAmutations detected in clinicalMtbisolates could confer FQ resistance, but several mutations reduced bacterial growth rates. Overall, recombinantM. smegmatisappears to be a beneficial surrogate system to evaluate FQ susceptibility of virulent mycobacteria.