Chang Chung-Yu, Lin Heng-Jia, Chang Lin-Li, Ma Ling, Siu L Kristopher, Tung Yi-Ching, Lu Po-Liang
1 Department of Microbiology and Immunology, School of Medicine, College of Medicine, Kaohsiung Medical University , Kaohsiung, Taiwan .
2 Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University , Kaohsiung, Taiwan .
Microb Drug Resist. 2017 Jan;23(1):98-106. doi: 10.1089/mdr.2015.0212. Epub 2016 May 5.
We analyzed the replicon types, sizes, and restriction fragment length polymorphism (RFLP) typing of plasmids carrying extended-spectrum β-lactamase (ESBL) genes in Klebsiella pneumoniae isolates from Taiwan. Fifty-one Escherichia coli transconjugant strains with plasmids from ESBL-producing K. pneumoniae from the Taiwan Surveillance of Antimicrobial Resistance III Program in 2002 were included. All the 51 plasmids carried a bla gene, the majority of which were bla (28/51 [54.9%]). Plasmids ranged in size from 126 to 241 kb by S1 nuclease digestion and subsequent pulsed-field gel electrophoresis, and the most common plasmid size (37.3%) was 161-170 kb. The most common replicon type of plasmids was incompatibility group (Inc)A/C (60.8%). The IncA/C plasmids all carried bla (bla), and some also carried bla (bla) genes. All 51 plasmids could be typed with PstI, and 27 (52.9%) belonged to 10 clusters. Thirty-eight of the 51 plasmids were typable with BamHI, and 21 plasmids (55.3%) fell into 7 clusters. Plasmids in the same cluster belonged to the same incompatibility group, with the exception of cluster C6. In conclusion, IncA/C plasmids are the main plasmid type responsible for the dissemination of ESBL genes of K. pneumoniae from Taiwan. RFLP with PstI possessed better discriminatory power than that with BamHI and PCR-based replicon typing for ESBL-carrying plasmids in K. pneumoniae in this study. Greater than 50% of plasmids fell into clusters, and >60% of cluster-classified plasmids were present in clonally unrelated isolates, indicating that horizontal transfer of plasmids plays an important role in the spread of ESBL genes.
我们分析了台湾地区肺炎克雷伯菌分离株中携带超广谱β-内酰胺酶(ESBL)基因的质粒的复制子类型、大小及限制性片段长度多态性(RFLP)分型。纳入了2002年台湾地区抗菌药物耐药性监测Ⅲ项目中51株携带产ESBL肺炎克雷伯菌质粒的大肠杆菌转接合子菌株。51个质粒均携带bla基因,其中大多数为bla(28/51 [54.9%])。经S1核酸酶消化及随后的脉冲场凝胶电泳分析,质粒大小范围为126至241 kb,最常见的质粒大小(37.3%)为161 - 170 kb。质粒最常见的复制子类型为不相容群(Inc)A/C(60.8%)。IncA/C质粒均携带bla(bla),部分还携带bla(bla)基因。51个质粒均可用PstI进行分型,27个(52.9%)属于10个簇。51个质粒中的38个可用BamHI进行分型,21个质粒(55.3%)归入7个簇。同一簇中的质粒属于同一不相容群,但C6簇除外。总之,IncA/C质粒是台湾地区肺炎克雷伯菌ESBL基因传播的主要质粒类型。本研究中,对于肺炎克雷伯菌携带ESBL的质粒,PstI - RFLP比BamHI - RFLP及基于PCR的复制子分型具有更好的鉴别能力。超过50%的质粒归入簇中,且>60%归入簇的质粒存在于克隆无关的分离株中,这表明质粒的水平转移在ESBL基因传播中起重要作用。