摘要
目的 对一株多药耐药肺炎克雷伯菌进行全基因组测序并对全部耐药基因进行分析.方法 2023年10月从一名脑梗死患者的尿液中常规方法分离肺炎克雷伯菌,进行鉴定和药敏试验,并通过16sRNA测序进行菌株确认,采用三代测序技术进行全基因组测序,通过生物信息学相关软件进行质量控制及组装,通过综合抗菌药物耐药性数据库对全部耐药基因进行分析.结果 从一位脑梗死男性患者尿液中分离出肺炎克雷伯菌,编号为hnyy23-147,对临床常用的内酰胺类、喹诺酮类、四环素类、氨基糖苷类、磺胺类抗菌药物均耐药,对多黏菌素敏感;全基因组测序显示菌株含有1个染色体和3个质粒,全部耐药基因分布于染色体2号、3号质粒上,耐药基因包括 aad A2b、blasHV-159、fos A6、qnrS1、sul1、sul2、dfr A14、rmtB、blaKPC-2、blaSHv-12,染色体突变导致的耐药包括acrR突变、ompK35突变、ompK36突变和gyrB突变.结论 通过全基因组测序的方法可及时了解多药耐药肺炎克雷伯菌全部耐药基因和分子流行病学规律,有助于加强医院感染控制和抗菌药物合理使用,对防止多药耐药菌传播具有重要意义.
Abstract
OBJECTIVE To sequence the whole genome of a multi-drug resistant strain of Klebsiella pneumoniae and analyze all drug resistance genes.METHODS In Oct.2023,a strain of Klebsiella pneumoniae was isolated from the urine of a patient with cerebral infarction by routine methods,identified and tested for drug sensitivity,then identified by VITEK-2 and confirmed by 16sRNA sequencing.The third-generation sequencing technology was used for whole genome sequencing.The quality control and assembly of the drug resistance genes were per-formed by bioinformatics related software,and the drug resistance function annotation platform was used to ana-lyze all resistance genes.RESULTS Klebsiella pneumoniae,No.hnyy23-147,was isolated from the urine of a male patient with cerebral infarction,and was resistant to clinically used antimicrobial drugs such as lactams,quinolones,tetracyclines,aminoglycosides,and sulfonamides,while was sensitive to polymyxin.Whole genome sequencing showed that the strain contained one chromosome and three plasmids,and all the drug resistance genes were distributed on chromosome and plasmids 2 and 3.Resistance genes included aad A2b,blasHv-159,fosA6,qnrS1,sul1,sul2,dfrA14,rmtB,blaKPC-2,blaSHv-12.Resistance caused by chromosomal mutations included acrR,ompK35,ompK36 and gyrB mutations.CONCLUSION The whole genome sequencing method can provide a timely understanding of all drug resistance genes and molecular epidemiology of multidrug-resistant Klebsiella pneumoniae,which can help to strengthen the control of nosocomial infection and the rational use of antibiotics,and is of great significance to preventing the spread of multidrug-resistant bacteria.
基金项目
军队后勤重点课题基金资助项目(AWS21J005)
NETL
NSTL
万方数据