目的:采用全基因组测序技术,研究7株对头孢他啶-阿维巴坦(CZA)耐药的CRKP菌株缺乏常见抗菌耐药基因(ARGs)的替代耐药机制。
结果:使用ARG数据库CARD和VF数据库筛选ARG和毒力因子(VF),分别,并使用BLAST+的基因组注释数据进行鉴定。6株为ST11序列型(STs),一个是ST2123。ST11菌株比ST2123菌株含有更多的ARG。所有七种菌株均携带多种具有外排介导的抗生素抗性的ARG,包括OQXA,OQXB,tet(A),qacedltal,CRP,H-NS,Kpn-E,F,G,H,acra,LptD,acrB,acrD,cpxA,mdtB,还有mdtC.在大多数菌株甚至所有菌株中鉴定了这些外排介导的ARG。全基因组测序显示,ST11菌株携带多个潜在的原种(PP),基因组岛(GI),以及整合和共轭元素,而ST2123菌株携带独立的PP和GI。
结论:全基因组测序分析显示,这7株缺乏常见ARGs的CZA耐药CRKP菌株表现出外排介导的抗生素耐药性相关ARGs。CRKP抵抗CZA的主要机制是抗生素失活。除了tet(A),未发现ARGs和与外排相关的验证实验.本研究结果为CRKP对CZA的抗性机制提供了新的可能,我们将在未来通过实验来验证这一结论。
OBJECTIVE: To investigate alternative resistance mechanisms among seven ceftazidime-avibactam (CZA)-resistant carbapenem-resistant Klebsiella pneumoniae (CRKP) strains lacking common antimicrobial resistance genes (ARGs) using whole genome sequencing.
RESULTS: ARG and virulence factors (VFs) were screened using the ARG database CARD and the VF database, respectively, and identified using genomic annotation data with BLAST+. Six strains were ST11 sequence types (STs), and one was ST2123. ST11 strains harbored more ARGs than the ST2123 strains. All seven strains carried multiple ARGs with efflux-mediated antibiotic resistance, including oqxA, oqxB, tet (A), qacEdltal, CRP, H-NS, Kpn-E, F, G, H, acrA, LptD, acrB, acrD, cpxA, mdtB, and mdtC. These efflux-mediated ARGs were identified in most strains and even all strains. Whole genome sequencing revealed that the ST11 strain carried multiple potential prophages, genomic islands, and integrative and conjugative elements, while the ST2123 strain carried an independent potential prophages and a genomic island.
CONCLUSIONS: Whole genome sequencing analysis revealed that these seven CZA-resistant CRKP strains lacking common ARGs exhibited efflux-mediated antibiotic resistance-associated ARGs. The main mechanism by which CRKP resists CZA is antibiotic inactivation. Except for tet (A), no ARGs and validation experiments related to efflux were found. This study\'s results provide a new possibility for the resistance mechanism of CRKP to CZA, and we will verify this conclusion through experiments in the future.