| Objective:1.To know the infection characteristics and drug resistance spectrum characteristics of Klebsiella Pneumoniae（KPN）and 123 strains of Carbapenem-resistant Klebsiella Pneumoniae（CRKP）in our hospital from January2015 to December 2019;2.To study the drug resistance mechanism of CRKP in our hospital and provide laboratory basis for guiding the rational use of antimicrobials in clinical practice.3.To analyze the gene homology of CRKP by the Multilocus sequence typing（MLST）in our hospital,and provide a theoretical basis for timely and effective prevention and control of horizontal transmission of CRKP.Methods:1.The WHONET 5.6 software was used to analyze the clinical data and drug resistance of 1,514 strains of KPN and 123 strains of carbapenem-resistant Klebsiella pneumoniae isolated clinically in the First Affiliated Hospital of Jinan University in the past five years;2.The phenotypic screening of carbapenem enzyme were detected by the modified carbapenem inactivation method（m CIM）and EDTA-carbapenem inactivation method（EDTA-carbapenem inactivation method,e CIM）or EDTA collaborative test;3.The four most common carbapenemase resistance genes of CRKP,bla KPC,bla VIM,bla IMP and bla NDM,were specifically amplified by PCR.The positive amplified products were sequenced and compared to determine the type of carbapenemase gene,and then compared with the phenotype test results;4.The 7 housekeeping genes of CRKP were specifically amplified by PCR.The amplified positive products were sequenced and compared on the Pasteur website to obtain the ST type,and the homology analysis was performed on the e BURST software.Results:1.From January 2015 to December 2019,a total of 1514 cases of KPN were isolated.In the past 5 years,the average separation rate of KPN in our hospital was9.44%,and the average detection rate of CRKP was 1.5%;2.The source of the 1514 cases of KPN detected in the past 5 years was sputum,with a total of 717 strains,accounting for 47.36%;the second was urine,with a total of 356（23.51%）detected;and a total of 250 strains were detected in blood（16.51%）;among the 123 CRKP strains,sputum specimens were the most sourced,75 strains were detected accounting for 60.97%;21 strains from urine specimens accounted for17.07%,and 15 strains from blood specimens accounted for 12.20%;3.The 1514 KPN strains detected from clinical departments in the past 5 years ranked among the top 4 departments are ICU with 270 strains accounting for 17.83%,followed by respiratory ward with 179 strains accounting for 11.82%,neurosurgery ward with 138 strains accounting for 9.11%,and neurologyward with 101 strains accounted for 6.67%;the 123 CRKP strains detected in clinical departments ranked among the top 4 departments in ICU,33 strains accounted for 26.83%,followed by respiratory ward 26 strains accounted for 21.14%,neurosurgery ward 20 strains accounted for 16.26%,and neurology ward 18 strains accounted for 14.63%;4.The resistance rates of KPN detected in our hospital from 2015 to 2019 to quinolones,cephalosporins,monocyclics,tetracyclines,carbapenems and sulfa drugs are all on the rise;The resistance rate of 123 strains of CRKP to cephalosporins,enzyme inhibitor complex preparations,carbapenems and other antibacterial drugs reached 100%,and theresistance rate to quinolones,monocyclics,minocyclinesand tetracycline has also reached more than 95%,and the drug resistance rate to aminoglycosides is slightly lower,but it is also close to more than 80%,and the drug resistance rate to tigecycline is the lowest 4.23%;5.The average age of 123 clinically isolates CRKP patients was 67 years old,and the average length of stay was 33 days.The results of the study showed that there were significant differences between the effective and ineffective of treatment in patient’s gender,combined underlying diseases,surgery,ventilator/intubation,and urinary tube/gastric tube retention,etc（P<0.05）;6.The m CIM test of 123 strains of CRKP was positive,and the positive rate was100%.Among them,the e CIM test and EDTA synergy test of 5 strains of CRKP were positive,and the positive rate was 4.1%;7.Among 123 strains of CRKP,118 strains detected the bla KPC gene,with a detection rate of 95.9%;5 strains detected the bla IMP gene,but no bla VIM and bla NDMgenes were detected,and one strain detected both bla KPC and bla IMPgenes.After electrophoresis positive,the sequencing results of bla IMP amplified products were all identified as IMP-4 subtype by BLAST comparison.8.The 123 CRKP were classified by MLST,and3 ST types were obtained.Among them,ST11 was the most with 117 strains,accounting for 95.1%;followed by ST76,with 5 strains,accounting for 4.1%;there were one strain of ST2193,accounting for 0.8%.Conclusions:1.From 2015 to 2019,the detection rate of KPN and CRKP in our hospital has increased year by year,and the detection rate of CRKP has increased significantly.The rate of resistance to multiple antibacterial drugs has shown a significant upward trend.The clinical practice should be further strengthened.2.The combined detection of m CIM test and EDTA collaborative test can effectively screen carbapenem-producing Klebsiella pneumoniae;3.The production of KPC-type carbapenemase and IMP-4 type metalloenzyme are the main mechanism of the resistance of Klebsiella pneumoniae to imipenem and meropenem in our hospital.4.The ST11 CRKP is widely spread in our hospital,ST2193 and ST11 are closely related,and the epidemiological monitoring needs to be strengthened. |
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