WS19.4 Molecular epidemiology of hot-spots of mutation in antimicrobial resistance loci of Pseudomonas aeruginosa isolates from cystic fibrosis airways

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• 作者：L. Greipel ; N. Cramer ; J. Klockgether ; M. Dorda ; S. Mielke ; P. Chouvarine ; L. Wiehlmann ; B. Tü ; mmler
• 刊名：Journal of Cystic Fibrosis
• 出版年：2015
• 出版时间：June 2015
• 年：2015
• 卷：14
• 期：supp_S1
• 页码：S35
• 全文大小：68 K

文摘

High-throughput genome sequencing of serial isolates from cystic fibrosis (CF) airways collected over a 30-year period uncovered targets of antipseudomonal chemotherapy as hot-spots of mutation. Sporadic mutations were overrepresented in the loci gyrA, rpoB, ampD, mexA, mexS, mexY, mexZ, parS, parR, amrZ, envZ, oprD, oprM, pagL, spuE, spuF, fusA1 and fusA2.

To reveal the frequency and spectrum of single nucleotide variations and frame-shift mutations in these loci in the contemporary P. aeruginosa population residing in CF airways, deep amplicon sequencing was performed with loci-spanning 405–1554 bp large PCR products amplified from DNA pooled from two strain collections. 305 isolates were obtained from 209 CF patients seen at 51 CF centers in Europe and USA and 56 isolates were retrieved from 38 CF patients after they had received early antipseudomonal chemotherapy of first lower airways colonization with P. aeruginosa.

The impact of missense, stop and frame-shift mutations on bacterial antimicrobial susceptibility phenotype was assessed by determination of MIC values against amikacin, aztreonam, cefepime, ceftazidime, ciprofloxacin, colistin, doripenem, fosfomycin, ghentamycin, imipenem, levofloxacin, meropenem, piperacillin, piperacillin/sulbactam, piperacillin/tazobactam and tobramycin.

The spectrum of SNPs and indels and their association in the 18 susceptibility loci with the antipseudomonal resistance phenotype will be presented at the meeting. Supported by DFG (SFB 900, A2) and BMBF (DZL at BREATH).