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The Study of the Mechanisms of Resistance in Carbapenem Resistance Acinetobacter Baumannii

Author: LiGuangRong
Tutor: LiuJinBo
School: Luzhou Medical College
Course: Internal Medicine
Keywords: Acinetobacter Baumannii SELDI-TOF-MS Resistance gene Cluster analysis Proteomics
CLC: R446.5
Type: Master's thesis
Year: 2013
Downloads: 82
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Abstract


Objective To analysis40strains of Acinetobacter baumanniiresistant spectrum Clinical isolated from our hospital, select the Carbapenemresistant stains, use Polymerase Chain Reaction(PCR)technology to detectCarbapenem resistant genes, clarifiy the main mechanism of Carbapenemresistance Acinetobacter baumannii. provide a scientific basis to guide clinicalrational use of antimicrobial agents, control and prevent the prevalence of drugresistant strains. adopt the method of proteomics differences, useSELDI-TOF-MS technology to detect Carbapenem resistant and sensitivestrains protein fingerprint, combine with clustering analysis, to explore theclinical value of finding Carbapenem resistant strains affinity of near and far.Methods1. Collect Acinetobacter baumannii strains isolated from January2011to December2012in our hospital.2. Identificate and detect the MIC of15kinds of antibacterial drugs by MicroScan WalkAway96automaticmicrobe/drug susceptibility testing system identification, Analysis the resistantspectrum.3. Use Polymerase Chain Reaction (PCR) technology to detectCarbapenem resistance gene OXA-23,OXA-24,OXA-58, IMP, VIM;electrophoresis the PCR amplification products, sequence the PCR positiveamplification. compare the results with the GenBank database.4. Extraction ofprotein from all40strains of Acinetobacter baumannii strains. Detectedbacteria protein fingerprint by SELDI-TOF-MS technique and Au protein chip. 5. Analysis the bacteria protein profiles using Biomarker Wizard software3.1.Screen the differentially expressed proteins from Carbapenem resistantAcinetobacter baumannii strains and sensitive. Using SPSS19.0Clusteringanalysis Carbapenem resistance of Acinetobacter baumannii, trace the affinityof clinical infection. Results1. A total of40strains of Acinetobacter baumanniiwere isolated by clinical specimens in our hospital.2. Analysis the Carbapenemresistance spectrum of40clinical isolated strains of Acinetobacter baumannii,obtain22strains of Acinetobacter baumannii to Carbapenem resistance, theresistant rate was55%. The22strains of Acinetobacter baumannii tocommonly used antimicrobial drug resistance rate were100%.3. In22strainsof Acinetobacter baumannii Carbapenem resistant gene，OXA-23positive were16strains, the positive rate was72.7%, OXA-58positive were5strains, thepositive rate was22.7%, VIM positive were7strains, the positive rate was31.8%.1strain with OXA-23, OXA-58and VIM gene, the positive rate was4.5%,3strains of both OXA-23and OXA-58gene, the positive rate was13.6%,7strains of both OXA-23and VIM gene, the positive rate was31.8%; theOXA-24and IMP gene were not detected. OXA-23, OXA-58, VIM geneamplification products were sequenced by the corresponding gene sequenceanalysis results compared with NCBI BLAST program homology were99%,99%and97-99%.4detected by SELDI-TOF-MS combined with Au chip in theAcinetobacter baumannii protein, get all of Acinetobacter baumannii proteinfingerprinting.Through the statistical analysis found that carbapenem resistantand sensitive to Acinetobacter baumannii strains expressed protein have significant difference（P<0.05）.5. Clustering analysis of antimicrobial drugresistance to Carbapenem Acinetobacter baumannii mass spectrometry results,the results showed carbapenem resistance of Acinetobacter baumannii ismainly in A type, B type of. Conclusion1. Acinetobacter baumanniicarbapenem antibacterial drug resistance rate was relatively high, especiallyImipenem was55%. And another14kinds of commonly used antimicrobialresistance rate was70%.2. The carbapenem resistant gene OXA-23was72.7%,carrying OXA-23gene may be mainly reason to carbapenem antibacterial drugresistant Acinetobacter baumannii, And one stain carrying OXA-23, OXA-58and VIM genes, three carrying OXA-23and OXA-58gene, seven carryingOXA-23and VIM genes, indicated that the important reason of Acinetobacterbaumannii resistant to antimicrobial agents commonly used in clinical wascarried multiple resistance.3.Carbapenem resistant Acinetobacter baumanniihad characteristic protein fingerprint. Carbapenem resistant Acinetobacterbaumannii distribution clustering analysis showed mainly type A, type B,A-type is divided into two subtypes A1and A2, and A1-based, type B is alsodivided into B1, B2, and B2-based.Carbapenem antibacterial drug resistantAcinetobacter baumannii protein profiling results clustering analysis candetermine the bacterial genetic affinity of distance, and can provide the basisfor clinical epidemiology detection.

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