Chen Wenyao, Fang Tingzi, Zhou Xiujuan, Zhang Daofeng, Shi Xianming, Shi Chunlei
Ministry of Science and Technology-United States Department of Agriculture Joint Research Center for Food Safety, School of Agriculture and Biology, Shanghai Jiao Tong University Shanghai, China.
Front Microbiol. 2016 Sep 30;7:1566. doi: 10.3389/fmicb.2016.01566. eCollection 2016.
The wide usage of antibiotics contributes to the increase in the prevalence of antibiotic-resistant . Plasmids play a critical role in horizontal transfer of antibiotic resistance markers in . This study aimed to screen and characterize plasmid profiles responsible for antibiotic resistance in and ultimately to clarify the molecular mechanism of transferable plasmid-mediated antibiotic resistance. A total of 226 isolates were examined for antimicrobial susceptibility by a disk diffusion method. Thirty-two isolates (14.2%) were resistant to at least one antibiotic. The presence of plasmid-mediated quinolone resistance (PMQR) genes and -lactamase genes were established by PCR amplification. PCR-based replicon typing revealed that these 32 isolates represented seven plasmid incompatibility groups (IncP, HI2, A/C, FIIs, FIA, FIB, and I1), and the IncHI2 (59.4%) was predominant. Antibiotic resistance markers located on plasmids were identified through plasmid curing. Fifteen phenotypic variants were obtained with the curing efficiency of 46.9% (15/32). The cured plasmids mainly belong to the HI2 incompatibility group. The elimination of IncHI2 plasmids correlated with the loss of -lactamase genes ( and ) and PMQR genes ( and ). Both IncHI2 and IncI1 plasmids in a serovar Indiana isolate SJTUF 10584 were lost by curing. The -carrying plasmid pS10584 from SJTUF 10584 was fully sequenced. Sequence analysis revealed that it possessed a plasmid scaffold typical for IncI1 plasmids with the unique genetic arrangement of IS-ΔIS----Δ inserted into the colicin gene . These data suggested that IncHI2 was the major plasmid lineage contributing to the dissemination of antibiotic resistance in and the activity of multiple mobile genetic elements may contribute to antibiotic resistance evolution and dissemination between different plasmid replicons.
抗生素的广泛使用导致了抗生素耐药性患病率的增加。质粒在细菌中抗生素耐药性标记的水平转移中起着关键作用。本研究旨在筛选和鉴定导致细菌抗生素耐药性的质粒图谱,并最终阐明可转移质粒介导的抗生素耐药性的分子机制。通过纸片扩散法对总共226株细菌分离株进行了药敏试验。32株分离株(14.2%)对至少一种抗生素耐药。通过PCR扩增确定了质粒介导的喹诺酮耐药(PMQR)基因和β-内酰胺酶基因的存在。基于PCR的复制子分型显示,这32株分离株代表了7个质粒不相容群(IncP、HI2、A/C、FIIs、FIA、FIB和I1),其中IncHI2(59.4%)占主导。通过质粒消除鉴定了位于质粒上的抗生素耐药性标记。获得了15个表型变体,消除效率为46.9%(15/32)。消除的质粒主要属于HI2不相容群。IncHI2质粒的消除与β-内酰胺酶基因(blaCTX-M-1和blaTEM-1)和PMQR基因(qnrS1和aac(6’)-Ib-cr)的丢失相关。一株印第安纳血清型分离株SJTUF 10584中的IncHI2和IncI1质粒通过消除而丢失。对来自SJTUF 10584的携带blaCTX-M-1的质粒pS10584进行了全序列测定。序列分析表明,它具有IncI1质粒典型的质粒支架,在大肠杆菌素基因colE1中插入了IS26-ΔIS26----ΔIS26的独特基因排列。这些数据表明,IncHI2是导致细菌中抗生素耐药性传播的主要质粒谱系,多种移动遗传元件的活性可能有助于抗生素耐药性在不同质粒复制子之间的进化和传播。