Study On The Genotype Of Klebsiella Pneumoniae Carbonase

Part? Resistance and homology analysis of carbapenem-resistant Klebsiella pneumoniae Objective To collect non-repetitive carbapenem-resistant Klebsiella pneumoniae isolated from our hospital,and the drug sensitivity test and homology were analyzed to understand the drug resistance of carbapenem-resistant Klebsiella pneumoniae(CRKP).To understand the drug resistance of carbapenem-resistant Klebsiella pneumoniae in our hospital,and to provide a reasonable experimental reference for clinical scientific use of drugs and prevention of CRKP infection.Method Klebsiella pneumoniae isolated from the First Affiliated Hospital of Anhui Medical University from January to December 2016 were collected and screened.Bacterial identification and susceptibility test were carried out with the full automatic microbiological analysis system of the French Mérieux VITEK 2 compact.K-B method was used as a supplementary drug susceptibility test and ERIC-PCR was used to analyze the homology of drug-resistant strains.Results A total of 25 strains of CRKP were collected in this study,which were mainly distributed in the ICU department,accounting for 32%,followed by Hematology departments,accounting for 20%,and infected specimens were mainly blood,accounting for 68%.The drug susceptibility test showed that the drug resistance rates of cefepime,ciprofloxacin,levofloxacin and furantoin were more than 90%,and all of them were resistant to piperacillin/tazobactam,cefuroxime,ceftazidime and aztreonam.The susceptibility rates to compound sulfamethoxazole and minocyclinewere 72% and 92%,respectively,and all of them were sensitive to polymyxin B.Twenty-five CRKP ERIC-PCR strains showed 8 genotypes: ????????????? and ?.Among them,II was the main epidemic strain,accounting for 56%.Conclusion The isolation of CRKP in our hospital was multi-drug resistant.It was difficult to prevent and treat Klebsiella pneumoniae,which posesed a great threat to clinical anti-infective treatment.We should strengthen prevention and standardize the use of antibiotics in clinic.ERIC-PCR showed that type II was an epidemic strain.Part? Detection of carbapenem-resistant Klebsiella pneumoniae genotype and retrospective analysis of clinical dataObjective The phenotype of CRKP was detected,and the efficacy of various tests were evaluated to provide a reasonable method for the detection of carbapenemase phenotype in clinical laboratories.The genotype of carbapenemase of Klebsiella pneumoniae was determined by PCR.The plasmid conjugation test was used to explore the horizontal transfer of drug resistance genes between strains.To clarify the genotype of carbapenemase in Klebsiella pneumoniae in our hospital,and the drug resistance can be transmitted between bacteria through plasmids and control the outbreak of drug-resistant strains in hospitals.By collecting clinical data of Klebsiella pneumoniae patients,retrospective analysis of the clinical characteristics of patients to explore the clinical related factors.Methods 25 strains of CRKP were screened and the carbapenemase phenotype was modified by modified Hodge test(MHT),Carba NP test and modified carbapenem inactivation method(m CIM).Imipenem-EDTA disk method was used to detect whether the strain produced metallo-?-lactamase.The two clinical datas of carbapenem-sensitive Klebsiella pneumoniae(CSKP)and CRKP were queried and analyzed.Result Twenty-five strains were tested by MHT test,Carba NP test and m CIM test.The results were consistent.The positive rate was 96%(24/25),and the negative strain was the same sample.The class B enzymes(metalloenzyme)was not detected in the synergistic test.The results of PCR sequencing confirmed that 24 strains of carbapenemase phenotype test were all KPC-2 gene,while the negative strains carried NDM-1 gene.Carbapenem-resistant genes were obtained from the conjugated bacteria,and their resistance to meropenem and imipenem increased by plasmid binding experiments.Statistical analysis of 25 patients with CRKP and 35 patients with CSKP showed that the proportion of blood samples,carbapenems and invasive treatment in CRKP group was significantly higher than that in CSKP group(P < 0.05).Conclusion Modified Hodge test,Carba NP test and m CIM test can accurately screen the carbapenemase-producing Klebsiella pneumoniae.K.pneumoniae carrying KPC-2and NDM-1 type carbapenemase genes were isolated in our hospital,these gens were located in plasmids,these gens could be transferred between strains through plasmids.Clinical datas show that invasive treatment and the use of carbapenems are the main factors leading to the production of carbapenem-resistant Klebsiella pneumoniae.