Sun Lingyan, Chen Yan, Qu Tingting, Shi Keren, Han Xinhong, Wu Wenhao, Jiang Yan, Yu Yunsong
Department of Laboratory Medicine, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, People's Republic of China.
Key Laboratory of Clinical in Vitro Diagnostic Techniques of Zhejiang Province, Hangzhou, People's Republic of China.
Infect Drug Resist. 2023 Dec 12;16:7621-7628. doi: 10.2147/IDR.S435195. eCollection 2023.
We aimed to characterize a novel and co-carrying hybrid plasmid from a clinical carbapenem-resistant (CRKP) strain.
Antimicrobial susceptibility was determined by the broth microdilution method. Plasmid size and localization were estimated using S1 nuclease pulsed-field gel electrophoresis (S1-PFGE) and Southern blotting. Plasmid transfer ability was evaluated by conjugation experiments. Whole genome sequencing (WGS) was performed using Illumina NovaSeq6000 and Oxford Nanopore MinION platforms. Genomic characteristics were analyzed using bioinformatics methods.
Strain ZY27320 was a multidrug-resistant (MDR) clinical ST11 strain that confers high-level resistance to carbapenems (meropenem, MIC 128 mg/L; imipenem, MIC 64 mg/L) and ceftazidime/avibactam (MIC >128/4 mg/L). Both S1-PFGE-Southern blotting and whole genome sequencing revealed that the carbapenemase genes and were carried by the same IncFII:IncR:IncN hybrid plasmid (pKPC2_NDM5). Conjugation experiments indicated that pKPC2_NDM5 was a non-conjugative plasmid.
This is the first report of a hybrid plasmid carrying both carbapenemase genes and in a clinical ST11 isolate that confers resistance to both ceftazidime/avibactam and carbapenems, thereby presenting a serious threat to treatment in clinical practice.
我们旨在鉴定一株临床耐碳青霉烯类肺炎克雷伯菌（CRKP）菌株携带的一种新型共携带杂交质粒。
采用肉汤微量稀释法测定抗菌药物敏感性。使用S1核酸酶脉冲场凝胶电泳（S1-PFGE）和Southern印迹法估计质粒大小和定位。通过接合实验评估质粒转移能力。使用Illumina NovaSeq6000和Oxford Nanopore MinION平台进行全基因组测序（WGS）。使用生物信息学方法分析基因组特征。
菌株ZY27320是一株多重耐药（MDR）临床ST11菌株，对碳青霉烯类药物（美罗培南，MIC 128 mg/L；亚胺培南，MIC 64 mg/L）和头孢他啶/阿维巴坦（MIC>128/4 mg/L）具有高水平耐药性。S1-PFGE-Southern印迹法和全基因组测序均显示，碳青霉烯酶基因由同一IncFII:IncR:IncN杂交质粒（pKPC2_NDM5）携带。接合实验表明pKPC2_NDM5是一种非接合性质粒。
这是首次报道在临床ST11分离株中存在同时携带碳青霉烯酶基因且对头孢他啶/阿维巴坦和碳青霉烯类药物均耐药的杂交质粒，从而对临床治疗构成严重威胁。
