环丙沙星诱导对临床耐药菌株主动外排基因转录的影响

doi:10.11843/j.issn.0366-6964.2017.09.022

Abstract

Abstract:

This study was conducted to explore the effect of ciprofloxacin on the transcription of several active efflux system related genes, acrA, acrB, acrD, acrE, acrF, mdtA, marA, robA and soxS,in clinical E. coli strains Y35 and J45.The minimal inhibitory concentrations (MICs) of ciprofloxacin to E. coli strains Y35 and J45 induced by ciprofloxacin were determined using the standard broth microdilution method. A real-time fluorescent quantitative PCR assay was established to observe the difference of active efflux pumps genes transcription level between parental strains and the 10^th, 20^th, 30^th generation of ciprofloxacin induced strains. Results were as follows:The results showed that, the 30^th generation of ciprofloxacin induced strains exhibited increased MICs of ciprofloxacin (2-fold), which was 256 μg·mL^-1. For Y35, 8 of 9 genes transcription level of the 10^th and 30^th of ciprofloxacin induced strains were increased compared with the parental strain, except gene mdtA, and the values of relative transcription level were between 1.20 and 96.07 (the differences were statistically significant, P<0.05 or P<0.01). The transcription level of acrD gene increased most markedly. The expression level of acrA, acrB and marA in the 20^th generation of ciprofloxacin induced strains were decreased compared with the 10^th generation strain, however,these genes showed increased transcription level compared with the parental strain. The relative transcription level of acrE, robA and soxS were between 1.40 and 3.81 (P<0.05 or P<0.01). For J45, only acrB and acrF gene transcription level were increased in the 10^th generation of ciprofloxacin induced strain, the relative value of transcription level were 2.76 and 1.73, however, the transcription level of other genes were decreased. acrA, acrB, acrE and mdtA gene transcription level were increased in the 20^th generation of ciprofloxacin induced strain, the value of relative transcription level were 15.35, 58.89, 31.56 36.50, respectively (P<0.01). All the genes transcription were increased to a high level in the 30^th generation of ciprofloxacin induced strain,the transcription level of acrF gene increased most markedly, it was 102.54 times of the parental strain. The results of this study suggest that continuous induction of the clinical resistance E. coli strains by subinhibitary concentration of ciprofloxacin might result in stronger resistance to ciprofloxacin, and increased level of transcription of active efflux pump genes. The increased transcription level of efflux pump genes might contribute to the stronger resistance of the induced strains to ciprofloxacin.

CLC Number:

S859.7

LIU Jian-hua, FU Sai-sai, WANG Ya-yun, HU Gong-zheng, PAN Yu-shan, YUAN Li. Transcription Changes of Active Efflux System in Clinical Multidrug-resistant E. coli Strains Induced by Ciprofloxacin Selective Pressure[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(9): 1753-1760.

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Transcription Changes of Active Efflux System in Clinical Multidrug-resistant E. coli Strains Induced by Ciprofloxacin Selective Pressure

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