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Study on Detection of AmpC Enzyme and Analysis of AmpC Gene in Enterobacter Cloacae

Author: HuYing
Tutor: ChenDuan
School: Kunming Medical College
Course: Clinical Laboratory Science
Keywords: Enterobacter cloacae AmpC enzyme Resistance AmpC gene ampD gene Mutation
CLC: R446.9
Type: Master's thesis
Year: 2005
Downloads: 55
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Abstract


Objective To understand the Kunming Enterobacter cloacae drug resistance and high of AmpC enzyme expression Enterobacter cloacae resistance . Explore the relationship between resistance and AmpC gene of Enterobacter cloacae Kunming . Methods KB ( disk diffusion ) analysis of drug resistance in clinical isolates of Enterobacter cloacae ; cefoxitin three-dimensional test , five pieces of paper phenotypic screening test, flucloxacillin dual inhibitor diffusion synergy test to detect high AmpC phenotypic expression ; regulate gene by polymerase chain reaction ( PCR ) assay structural gene ampC and ampD line on the part of the drug-resistant strains DNA sequencing to detect gene mutations . 74 Enterobacter cloacae except imipenem and cefepime higher sensitivity ( 98.6% and 86.5% ) , low sensitive to other antimicrobial agents (63.5% ) . Are three ways to detect high AmpC were 28.4% (21/ 74 ) , 35.1% ( 26 /74) , 37.8 % (28 /74) , showed no significant difference ( P GT ; 0.05 ) High AmpC strains and enzyme production strains resistant rate has a significant difference ( P lt ; 0.05 ) . PCR amplification of ampC and ampD gene positive rates were 89.2% and 86.5% , respectively . 4 of phenotypic strains for the high-yielding the AmpC enzymes strains ampC gene amplification product sequences with standard strains ampCP99 gene sequence comparison analysis , were 99% nucleotide homology , the deduced amino acid sequence 58 from alanine - proline ( Ala - 58 → Pro ) point mutation , the rest are synonymous mutations . The ampD gene nucleotide and the standard strain E.cloacae14ampD gene sequence comparison analysis revealed the presence of mutations , the corresponding sites of amino acid mutations Phe - 63 → Tyr , Ala - 68 → Ser , Met - 101 → Lys . Conclusion Kunming Enterobacter cloacae serious drug resistance , high AmpC may result in clinical isolates of Enterobacter cloacae main reason for antibiotic resistance . The five kinds of paper phenotypic screening tests and flucloxacillin dual inhibitor diffusion synergy test can be used as a routine screening of clinical microbiology laboratories AmpC enzyme . AmpC gene are widely distributed in Enterobacter cloacae ampD gene mutation may be associated with high AmpC .

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