Department of Clinical Laboratory, Zhejiang Hospital, Hangzhou, People's Republic of China.
State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, National Medical Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, People's Republic of China.
J Glob Antimicrob Resist. 2023 Dec;35:181-189. doi: 10.1016/j.jgar.2023.08.021. Epub 2023 Sep 20.
To characterize the evolution and interspecies transfer of plasmids between Klebsiella pneumoniae and Escherichia coli within a single patient.
Minimum inhibitory concentrations were measured using broth microdilution assays. Conjugation assays, string tests, and Galleria mellonella infection model experiments were also conducted. Whole-genome sequencing was performed on the Illumina and Nanopore platforms. Antimicrobial resistance determinants, insertion sequences, and virulence factors were identified using ABRicate/ResFinder database, ISFinder, and virulence factor database. Wzi and capsular polysaccharide (KL) were typed using Kleborate and Kaptive. Multi-locus sequence typing (MLST), replicon typing, and single nucleotide polymorphism analyses were conducted using the BacWGSTdb server.
The carbapenem-resistant K. pneumoniae 2111KP was characterized as ST11, wzi64, and KL64, with a positive string test result and a relatively high virulence phenotype. Analysis of the 2111KP genome revealed that bla was located in a 268,400-bp IncFIB/IncHI1B/IncX3 conjugative plasmid (p2111KP-1), regulated by IS26, IS5, and ISKox3. p2111KP-1 was also a rmpA2-associated virulence plasmid with an iutA-iucABCD gene cluster and a IS26-mediated multidrug-resistant fusion plasmid, which contained 8-bp (AGCTGCAC or GGCCTTTG) target site duplications. Segments flanked by IS26 of p2111KP-1 were 99.99% identical to a 49,016-bp E. coli plasmid.
This study provided direct evidence of plasmid fusion via IS26 between two different bacterial species within one patient and revealed the process by which genetic elements conferring carbapenem resistance and virulence were simultaneously transferred between these species. It highlights the need for strategic antibiotic use and rigorous monitoring to prevent the plasmid-mediated fusion and transmission of drug-resistance/virulence factors.
描述同一患者体内肺炎克雷伯菌和大肠杆菌之间质粒的进化和种间转移。
采用肉汤微量稀释法测定最小抑菌浓度。还进行了接合试验、串珠试验和大蜡螟感染模型实验。使用 Illumina 和 Nanopore 平台进行全基因组测序。使用 ABRicate/ResFinder 数据库、ISFinder 和毒力因子数据库鉴定抗生素耐药决定因子、插入序列和毒力因子。使用 Kleborate 和 Kaptive 对 Wzi 和荚膜多糖（KL）进行分型。使用 BacWGSTdb 服务器进行多位点序列分型（MLST）、复制子分型和单核苷酸多态性分析。
耐碳青霉烯肺炎克雷伯菌 2111KP 被鉴定为 ST11、wzi64 和 KL64，具有阳性串珠试验结果和较高的毒力表型。2111KP 基因组分析表明，bla 位于 268400bp IncFIB/IncHI1B/IncX3 可接合质粒（p2111KP-1）中，由 IS26、IS5 和 ISKox3 调控。p2111KP-1 也是一个 rmpA2 相关毒力质粒，带有 iutA-iucABCD 基因簇和一个由 IS26 介导的多药耐药融合质粒，其中包含 8bp（AGCTGCAC 或 GGCCTTTG）靶序列重复。p2111KP-1 中由 IS26 侧翼的片段与 49016bp 的大肠杆菌质粒 99.99%相同。
本研究提供了同一患者体内两种不同细菌种间质粒融合的直接证据，并揭示了携带碳青霉烯耐药和毒力基因的遗传元件在这些种间同时转移的过程。它强调了需要战略性使用抗生素并严格监测，以防止质粒介导的融合和耐药/毒力因子的传播。
