| Objective:To verify the accuracy of the improved Hodge experiment,to understand the distribution and composition of the bacterial resistance gene in 5 hospitals in Qingdao urban area,and to provide a basis for the rational application of antibiotic therapy in clinic by the screening experiment of carbapenemase phenotype and drug resistance gene detection experiment of carbapenem-resistant Klebsiella pneumoniae(CRKP)in Qingdao city.To explore the prospects and current status of clinically effective drugs,according to the related research of CRKP drugs worldwide.Method:This study collected a total of 54 CRKP infection specimens or CRKP strains that have been clinically confirmed in five top three hospitals in Qingdao city from October 2016 to December 2019;at the same time,selected non-tolerance clinically confirmed by the same hospital in the same time period A total of 55 Klebsiella pneumoniae strains of carbapenem antibiotics were used as the control group for drug sensitivity tests.The Meriere VITEK 2 Compact fully automatic microbial biochemical identification system was used for the identification of the strains for verification.The paper-sheet diffusion method was used to sequentially detect the tested strains against amikacin,compound neonomine,gentamicin,levofloxacin,imipenem-cilastatin sodium,piperacillin,tazobactam,meropenem,Drug resistance of cefotaxime and cefoperazone and sulbactam sodium,according to the diameter of the bacteriostatic ring,according to the CLSI M100-28 th drug susceptibility test performance standards for interpretation of drug resistance;modified Hodge test for carbapenemase phenotype Screening,the negative control was Klebsiella pneumoniae ATCC 700603 strain.According to the degree of invasion of the test strain on the E.coli quality control bacteria ATCC25922 medium,the degree of invasion of the bacteriostatic ring was judged;Finally,the boiling method is used to extract the DNA,and the extracted DNA is polymerized chain reaction(Polymerase chain reaction,PCR)to amplify the target genes of blaKPC-2,blaNDM-1,blaOXA-48,blaIMP,blaVIM,and agar the PCR products In the sugar gel electrophoresis test,the stained agarose gel is put into a multifunctional imaging system to observe and record the experiment results.Results:The collected CRKP strains have the largest source of respiratory sputum,accounting for 74.07%,and the infected elderly account for 85.19%.The number of patients in theintensive care department and the respiratory department is larger,accounting for 46.20%and 25.93%,respectively..The tested strains were verified by Merrier’s VITEK 2 Compact automatic microbial biochemical identification system,and the collected strains were Klebsiella pneumoniae.By interpreting the diameter of the bacteriostatic ring,the susceptibility test in this study showe that the collected strains were resistant to one or more carbapenem antibiotics,and the carbapenem antibiotic-resistant Klebsiella pneumoniae tested Generally has a high resistance rate to commonly used antibiotics in clinical practice,with the lowest resistance rate of amikacin,which is 35.2%;followed by compound sinomine 53.7%;gentamicin 55.6%;levofloxacin 75.9%;imipenem 88.9%;piperacillin-tazobactam 92.6%;cefotaxime and meropenem 94.4%;cefoperazone-sulbactam sodium,98.1%.;Improved Hodge experiment showed that among the 54 samples tested,43 were positive strains and 11 were negative strains,the positive rate was 79.6%;the final drug resistance gene test experiment showed that a total of carbapenems were detected There were 40 strains of enzyme resistance genes,and the detection rate of target resistance genes was 74.07%.Among them,35 strains were found to carry the KPC-2 gene,7 strains were to carry the OXA-48 gene,4 strains were to carry the NDM-1 gene,and 1 strain was to carry the IMP gene.All strains carrying the OXA-48 gene also carried the KPC-2 gene.Another 14 strains did not detect the target carbapenem gene.Conclusion:(1)The main infection site of the CRKP strain is the respiratory tract,the main distribution department is the Department of Intensive Medicine,and the main infection group is the elderly patients.(2)In addition to the compound Xinnuoming,the resistance of CRKP to the other 8antibiotics participating in the drug susceptibility test was significantly higher than that of non-CRKP strains,and the difference was statistically significant(P<0.05).(3)There is a false positive rate in the modified Hodge test.Although some strains do not produce carbapenemase,they may produce high-efficiency plasmid-mediated cephalosporinase,which still shows resistance to carbapenem antibiotics.(4)The carbapenem-resistant gene of carbapenem-resistant Klebsiella pneumoniae carried in Qingdao city was mainly KPC-2,followed by OXA-48,NDM-1 and IMP genes,and no VIM gene was detected..The drug resistance genes and CRKP detected in this experiment on amikacin,co-trimoxazole,gentamicin,levofloxacin,piperacillin-tazobactam,cefotaxime,cefoperazone-sulbactam sodium There is no obvious correlation for high drug resistance... |
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