铜绿假单胞菌外毒素U和外毒素S中消毒剂抗性基因与对多用途消毒溶液和抗生素敏感性之间的关联
Association between disinfectant resistance genes in exoU and exoS Pseudomonas aeruginosa with sensitivity to multipurpose disinfecting solutions and antibiotics.
作者信息
Akter Tanzina, Stapleton Fiona, Green Matthew, Willcox Mark
机构信息
School of Optometry and Vision Science, Faculty of Medicine and Health, University of New South Wales (UNSW), Sydney NSW-2052, Australia; Microbial Biotechnology Division, National Institute of Biotechnology (NIB), Dhaka-1349, Bangladesh.
School of Optometry and Vision Science, Faculty of Medicine and Health, University of New South Wales (UNSW), Sydney NSW-2052, Australia.
出版信息
Cont Lens Anterior Eye. 2025 May 13:102436. doi: 10.1016/j.clae.2025.102436.
BACKGROUND
The aim of this study was to determine differences in susceptibility of exoU and exoS Pseudomonas aeruginosa to multi-purpose disinfecting solutions (MPDS) and antibiotics and their association with disinfectant resistance genes.
METHODS
A total of 16 exoU and 24 exoS P. aeruginosa keratitis isolates were tested for susceptibility against four multipurpose disinfecting solutions (OPTI-FREE PureMoist, Acuvue RevitaLens, Biotrue, and Renu Advanced Formula) using minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations. Antibiotic susceptibility to ciprofloxacin, levofloxacin, gentamicin, and tobramycin was determined by MIC. Whole genome sequences from 27 isolates were utilized to identify disinfectant resistance genes and single nucleotide polymorphisms (SNPs) in the genes using the Comprehensive Antibiotic Resistance Database (CARD) and Geneious Prime respectively. Polymerase chain reaction (PCR) was used to detect the disinfectant resistance genes qacE, qacEΔ1, and sugE1. Correlation between disinfectant resistance genes and their SNPs with disinfectant and antibiotic MICs was determined by Spearman's Rho.
RESULTS
All MPDS were bactericidal at full strength, but upon dilution OPTI-FREE Puremoist and Renu Advanced Formula were the most active against the exoU and exoS groups correspondingly. ExoU showed significantly higher resistance than exoS (p ≤ 0.04) to levofloxacin (37.5 % vs. 8.3 %), gentamicin (43.8 % vs. 0 %), and tobramycin (37.5 % vs. 0 %). The qacEΔ1 and sugE1 genes along with several SNPs in the resistance genes were more common in the exoU group. SNPs in disinfectant resistance genes correlated with MPDS MICs/MBCs, notably mexQ Arg1036His (R ≥ -0.38, p ≤ 0.04), Gly505Asp (R ≥ 0.40, p ≤ 0.04) and opmD Gly269Ser (R ≥ 0.43, p ≤ 0.03). QacEΔ1 and sugE1 were linked to increased MIC (R ≥ 0.54, p < 0.01) and MBC (R ≥ 0.50, p < 0.01) for Renu Advanced Formula. Also, multiple SNPs and qacEΔ1 and sugE1 showed moderate to large positive associations with antibiotic resistance.
CONCLUSIONS
ExoU strains had distinct MPDS susceptibility patterns, with higher disinfectant resistance genes prevalence and mutations, contributing to MPDS and antibiotics resistance.
背景
本研究旨在确定外排U型和外排S型铜绿假单胞菌对多用途消毒溶液（MPDS）和抗生素的敏感性差异，以及它们与消毒剂抗性基因的关联。
方法
使用最低抑菌浓度（MIC）和最低杀菌浓度（MBC），对总共16株外排U型和24株外排S型铜绿假单胞菌角膜炎分离株进行了针对四种多用途消毒溶液（OPTI-FREE PureMoist、Acuvue RevitaLens、Biotrue和Renu Advanced Formula）的敏感性测试。通过MIC确定对环丙沙星、左氧氟沙星、庆大霉素和妥布霉素的抗生素敏感性。利用来自27株分离株的全基因组序列，分别使用综合抗生素抗性数据库（CARD）和Geneious Prime来鉴定消毒剂抗性基因和这些基因中的单核苷酸多态性（SNP）。采用聚合酶链反应（PCR）检测消毒剂抗性基因qacE、qacEΔ1和sugE1。通过Spearman秩相关分析确定消毒剂抗性基因及其SNP与消毒剂和抗生素MIC之间的相关性。
结果
所有MPDS在原液浓度下均具有杀菌作用，但稀释后，OPTI-FREE Puremoist和Renu Advanced Formula分别对外排U型和外排S型组最为有效。外排U型对左氧氟沙星（37.5%对8.3%）、庆大霉素（43.8%对0%）和妥布霉素（37.5%对0%）的耐药性显著高于外排S型（p≤0.04）。qacEΔ1和sugE1基因以及抗性基因中的几个SNP在外排U型组中更为常见。消毒剂抗性基因中的SNP与MPDS的MIC/MBC相关，特别是mexQ Arg1036His（R≥-0.38，p≤0.04）、Gly505Asp（R≥0.40，p≤0.04）和opmD Gly269Ser（R≥0.43，p≤0.03）。QacEΔ1和sugE1与Renu Advanced Formula的MIC（R≥0.54，p<0.01）和MBC（R≥0.50，p<0.01）升高有关。此外，多个SNP以及qacEΔ1和sugE1与抗生素耐药性呈中度至高度正相关。
结论
外排U型菌株具有独特的MPDS敏感性模式，消毒剂抗性基因的流行率和突变较高，导致对MPDS和抗生素产生耐药性。
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