Department of Pharmacy, Shaoxing Hospital of Traditional Chinese Medicine Affiliated to Zhejiang Chinese Medical University, Shaoxing, China.
Department of Pharmacy, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Front Cell Infect Microbiol. 2022 Sep 29;12:984479. doi: 10.3389/fcimb.2022.984479. eCollection 2022.
To characterize one OXA-232-producing -KL112-O1 carbapenem-resistant (CRKP) co-harboring chromosomal and one -associated virulence plasmid.
Minimum inhibitory concentrations (MICs) were measured broth microdilution method. Conjugation, chemical transformation, string test and infection model experiments were also conducted. Whole-genome sequencing (WGS) was performed on the Illumina and Nanopore platforms. Antimicrobial resistance determinants were identified using ABRicate program with ResFinder database. Insertion sequences (ISs) were identified using ISfinder. Bacterial virulence factors were identified using virulence factor database (VFDB). Wzi, capsular polysaccharide (KL) and lipoolygosaccharide (OCL) were analyzed using Kleborate with . Phylogenetic analysis of 109 ST15 strains was performed using core genome multilocus sequence typing (cgMLST) on the Ridom SeqSphere+ server. MLST, replicons type, SNP strategies and another cgMLST analysis for 45 OXA-232-producing strains were further conducted using BacWGSTdb server.
KPTCM strain belongs to ST15 with , KL112 and O1. It possessed a multidrug-resistant (MDR) profile and was resistant to carbapenems (meropenem and ertapenem), ciprofloxacin and amikacin. Virulence assays demonstrated KPTCM strain possesses a low virulence phenotype. WGS revealed it contained one circular chromosome and nine plasmids. The carbapenemase-encoding gene was located in a 6141-bp ColKP3-type non-conjugative plasmid and flanked by ΔIS and Δ-Δ. Interestingly, was located in the chromosome mediated by ISbased transposon Tn. Importantly, it harbored a -associated pLVPK-like virulence plasmid with - gene cluster and one IS-mediated MDR fusion plasmid according to 8-bp (AGCTGCAC or GGCCTTTG) target site duplications (TSD). Based on the cgMLST and SNP analysis, data showed OXA-232-producing ST15 isolates were mainly isolated from China and have evolved in recent years.
Early detection of CRKP strains carrying chromosomal , OXA-232 carbapenemase and pLVPK-like virulence plasmid is recommended to avoid the extensive spread of this high-risk clone.
对一株同时携带染色体型 OXA-232 型碳青霉烯酶和 1 个 -相关毒力质粒的 -KL112-O1 型碳青霉烯酶耐药 （CRKP） 肺炎克雷伯菌进行特征分析。
采用肉汤微量稀释法测定最小抑菌浓度 （MIC）。进行了接合、化学转化、串联试验和感染模型实验。使用 Illumina 和 Nanopore 平台进行全基因组测序 （WGS）。使用 ABRicate 程序和 ResFinder 数据库鉴定抗生素耐药决定因子。使用 ISfinder 鉴定插入序列 （IS）。使用毒力因子数据库 （VFDB） 鉴定细菌毒力因子。使用 Kleborate 分析 Wzi、荚膜多糖 （KL） 和脂寡糖 （OCL）。在 Ridom SeqSphere+ 服务器上使用核心基因组多位点序列分型 （cgMLST） 对 109 株 ST15 菌株进行系统发育分析。使用 BacWGSTdb 服务器对 45 株产 OXA-232 的 菌株进行 MLST、复制子类型、SNP 策略和另一个 cgMLST 分析。
KPTCM 株属于 ST15，带有 KL112 和 O1。它具有多药耐药 （MDR） 表型，对碳青霉烯类 （美罗培南和厄他培南）、环丙沙星和阿米卡星耐药。毒力试验表明 KPTCM 株具有低毒力表型。WGS 显示它含有一个圆形染色体和九个质粒。碳青霉烯酶编码基因 位于 6141-bp ColKP3 型非接合性质粒中，两侧为 ΔIS 和 Δ-Δ。有趣的是， 位于由 IS 介导的转座子 Tn 介导的染色体上。重要的是，根据 8 个碱基对 （AGCTGCAC 或 GGCCTTTG） 靶位重复 （TSD），它携带一个 -相关的 pLVPK 样毒力质粒，其中包含 - 基因簇和一个由 IS 介导的 MDR 融合质粒。根据 cgMLST 和 SNP 分析，数据显示产 OXA-232 的 ST15 分离株主要分离自中国，并且是近年来进化而来的。
建议早期检测携带染色体型 OXA-232 型碳青霉烯酶和 pLVPK 样毒力质粒的 CRKP 菌株，以避免这种高风险克隆的广泛传播。
