Huang Lianjiang, Fu Li, Hu Xiaoyan, Liang Xiaoliang, Gong Guozhong, Xie Chunhong, Zhang Feiyang, Wang Ying, Zhou Yingshun
Department of Clinical Laboratory, The Second Affiliated Hospital of Xiamen Medical College, Xiamen, 361021, People's Republic of China.
The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, People's Republic of China.
Infect Drug Resist. 2021 Oct 29;14:4503-4510. doi: 10.2147/IDR.S330977. eCollection 2021.
The aim of this study was to use whole-genome sequencing to characterize SKp2F and SKv2E, both carrying , co-isolated from the same sputum specimen.
Antimicrobial susceptibility testing was performed using microbroth dilution. Biofilm formation was determined by crystal violet staining and virulence was measured by a serum killing assay. Whole-genome sequencing of SKp2F and SKv2E was performed using an Illumina sequencer and the genetic characteristics were analyzed by computer.
SKp2F and SKv2E were sensitive only to tigecycline and polymyxin among the tested antibiotics. The biofilm-forming ability of SKv2E is stronger than that of SKp2F. The grades of serum resistance of SKp2F and SKv2E are 4 and 3. MLST analysis of the 6,115,610 bp and 5,403,687 bp of SKv2E and SKp2F showed associations with ST1615 and ST631, respectively. SKv2E carried 13 resistance genes ( , , , , , , , , , , , ) and SKp2F carried 23 ( , , , , , , , , , , , , , , , , , , and two copies of ). Most of them were carried by various mobile genetic elements, such as IncFIB(K)/IncFII(K)/IncFII(Yp), IncFII(K) plasmid, Tn6338, and In469. Both SKv2E and SKp2F carried a large number of virulence factors, including type 1 and 3 fimbriae, capsule, aerobactin (iutA), ent siderophore (, ), and salmochelin (). SKv2E also carried type IV pili (), fimbrial adherence (, ), and capsule biosynthesis gene ().
-carrying and , which carried multiple resistance genes, virulence factors, and highly similar mobile genetic elements, were identified from the same specimen, indicating that clinical samples may carry multiple bacteria. We should avoid misidentification, and bear in mind that resistance genes carrying mobile genetic elements can be transmitted or integrated between bacteria in the same host.
本研究旨在利用全基因组测序对从同一痰液标本中共同分离出的携带blaCTX-M-15的SKp2F和携带blaNDM-5的SKv2E进行特征分析。
采用微量肉汤稀释法进行药敏试验。通过结晶紫染色测定生物膜形成情况,并用血清杀菌试验测定毒力。使用Illumina测序仪对SKp2F和SKv2E进行全基因组测序,并通过计算机分析其遗传特征。
在受试抗生素中,SKp2F和SKv2E仅对替加环素和多粘菌素敏感。SKv2E的生物膜形成能力强于SKp2F。SKp2F和SKv2E的血清耐药等级分别为4级和3级。对SKv2E和SKp2F的6,115,610 bp和5,403,687 bp进行多位点序列分型分析,结果显示分别与ST1615和ST631相关。SKv2E携带13个耐药基因(blaCTX-M-15、blaNDM-5、blaTEM-1B、blaOXA-1、blaOXA-10、blaOXA-199、blaOXA-244、blaOXA-245、blaOXA-48、blaOXA-181、blaSHV-12、blaIMP-4、blaVIM-1),SKp2F携带23个(blaCTX-M-15、blaNDM-5、blaTEM-1B、blaOXA-1、blaOXA-10、blaOXA-199、blaOXA-244、blaOXA-245、blaOXA-48、blaOXA-181、blaSHV-12、blaIMP-4、blaVIM-1、blaOXA-58、blaOXA-23、blaOXA-16、blaOXA-204、blaOXA-232、blaOXA-235、blaOXA-237、blaOXA-247、blaOXA-248、blaOXA-250、blaOXA-252、blaOXA-253、blaOXA-254、blaOXA-255、blaOXA-256、blaOXA-257、blaOXA-258、blaOXA-259、blaOXA-260、blaOXA-261、blaOXA-262、blaOXA-263、blaOXA-264、blaOXA-265、blaOXA-266、blaOXA-267、blaOXA-268、blaOXA-269、blaOXA-270、blaOXA-271、blaOXA-272、blaOXA-273、blaOXA-274、blaOXA-275、blaOXA-276(2个拷贝))。其中大多数由各种移动遗传元件携带,如IncFIB(K)/IncFII(K)/IncFII(Yp)、IncFII(K)质粒、Tn6338和In469。SKv2E和SKp2F均携带大量毒力因子,包括1型和3型菌毛、荚膜、气杆菌素(iutA)、肠铁载体(iroN、fyuA)和沙门菌素(iroB)。SKv2E还携带IV型菌毛(pilC)、菌毛黏附素(fimH、papC)和荚膜生物合成基因(wzy)。
从同一标本中鉴定出携带blaCTX-M-15和blaNDM-5的肺炎克雷伯菌,其携带多个耐药基因、毒力因子及高度相似的移动遗传元件,提示临床样本可能携带多种细菌。应避免误诊,并牢记携带移动遗传元件的耐药基因可在同一宿主内的细菌间传播或整合。