| Objective:1 To grasp the situation and characteristics of drug resistance of Pseudomonas aeruginosa in Baoding,in order to guide clinicians rational using antibiotics,slow down antibiotic producing selection pressure to bacterium drug-resistant,delay the state of bacterium drug-resistantand and control the hospital infection.2 To select imipenem-resistant pseudomonas aeruginosa,detect the type of ?-lactamasese gene and the Opr D2 gene deletion status,to explore the mechanism of drug resistance to imipenem-resistant Pseudomonas Aeruginosa.Methods:1 The specimen sourceSamples were collected from the Pseudomonas aeruginosa isolated from the hospital patients from 2016.7 to 2017.1 in the No.1center hospital in baoding.According to The Ministry of Public Health National Guide to Clinical Laboratory Procedures(Third Edition),Pseudomonas aeruginosa were isolated from clinical specimens by Sheep blood agar,and identificated by Using VITEK-2,which from the French bio Merieux company.2 The drug sensitivity testThe sensitivities of 23 antibiotics were detected using automatic bioanalys VITEK-2 and K-B method in the pseudomonas aeruginosa strains isolated after purification,and select the imipenem-resistant Pseudomonas aeruginosa.3 The detection of drug resistance genesThe DNA was extracted using Maxwell?16 Cell LEV DNA Purification Kit,the ?-lactamasese gene and the Opr D2 gene were amplified using polymerase chain reaction(PCR)in imipenem-resistant strains.The product was conducted electrophoresis in 2% agarose gel,the results of electrophoresis was observed by gel imagerand and compared to DNAMarker of the standard amount.The results of electrophoresis of PCR products were analyzed and photograhed.,with the same gene fragment was regarded as positive bands and the photograph was preserved by camera.Results:1 Drug resistance of Pseudomonas aeruginosaThe drug-resistance rate of 122 Pseudomonas aeruginosa strains isolated from 2016.7 to 2017.1 were respective to the 23 kinds of antibiotics as following: ampicillin98.36%,amikacin10.66%,ciprofloxacin25.41%,gentamicin16.39%,nebcin15.57%,cotrimoxazole96.72%,ampicillin/sulbactam96.72%,ceftazidime17.21%,imipenem40.98%,piperacillin18.03%,cefazolin97.54%,cefepime20.49%,levofloxacin22.95%,meropenem23.77%,cefotetan95.9%,piperacillin and tazobactam(conventionaldosage)13.11%,efatriaxone98.36%,cefoperazone36.07%,efoperazone/sulbactam15.57%,cefuroximeester95.9%,ctefuroximesodium95.9%,biapenem23.77%,the furadantin was only used to the urine,and the drug-resistance rate is 100%.2 The detection of ?-lactamases gene detectionThe positive of ?-lactamases among the 50 imipenem-resistant Pseudomon asaeruginosa:TEM(3stains),PER(3stains),VEB(1stain),GES(2stains),OX A-2(2stains),OXA-10(3stains),CTX-M-1(3stains).3 The detection of Metallo-?-lactamase gene detection The positive of Metallo-?-lactamase among the 50 imipenem-resistant Pseudomonas aeruginosa: VIM(9stains),IMP(11stains),SIM(0stains),SPM(5stains),GIM(0stain).4 The missing of Membrane protein Opr D2Among the 50 imipenem-resistant Pseudomonas aeruginosa,there are 31 strains Opr D2 missing,missing 62%.Conclusion:1 The rate of drug-resistant is high in cefuroxime ester,ctefuroxime sodium,cefotetan,efatriaxone,ampicillin,cotrimoxazole in baoding distract.2 The drug-resistance rate of Pseudomonas aeruginosa to imipenem is40.98%.3 Among the 50 imipenem-resistant Pseudomonas aeruginosa,the detective rate of ?-lactamases gene is 58%(29stains),and containing there are10 strains containing 2 or more than 2 genes;and the loss rate of Opr D2 is high,missing 62%,which point out that the resistant of Pseudomonas aeruginosa to ?-lactamases is controlled by many kinds of drug-resistant genes together,and have complex drug-resistant mechanism.4 We should strengthen the monitoring of antibiotic resistance of Pseudomonas aeruginosa and the study on molecular epidemiology of resistance genes,in order to prevent the spread of drug-resistant strains,and improve the clinical cure rate. |
PDF Full Text Request