The Relation between the High-level Resistance to Fluoroquinolones and the Over-expression of the acrA among Quinolone-Resistant Escherichia coli
- Authors
- 황병연; 기세윤; 김정연; 정혜원; 박정원; 박윤희; 김명곤; 정희진; 김우주
- Issue Date
- Aug-2005
- Publisher
- Korean Society of Infectious Diseases; Korean Society for Antimicrobial Therapy
- Keywords
- AcrA; quinolone resistant E. coli; efflux pump; Real-time PCR; Northern hybridization
- Citation
- Infection and Chemotherapy, v.37, no.4, pp 185 - 192
- Pages
- 8
- Indexed
- KCICANDI
- Journal Title
- Infection and Chemotherapy
- Volume
- 37
- Number
- 4
- Start Page
- 185
- End Page
- 192
- URI
- https://scholarworks.korea.ac.kr/kumedicine/handle/2020.sw.kumedicine/19970
- ISSN
- 2093-2340
2092-6448
- Abstract
- Background
Target point mutation of DNA topoisomerase, which is the typical mode of quinolone resistance, cannot explain high level resistance to quinolones. Therefore, many authors looked into over expression of efflux pump as the possibility. After quantificating the arcA mRNA, which controls AcrABTolC, the authors tried to find out the difference in the expression of arcA mRNA according to MIC of ciprofloxacin. The authors also tried to determine the usefulness of real time PCR, which is more reproducible and takes less time than preexisting immunoblot assay, through quantification of acrA.
Materials and Methods
Mutations in topoisomerase (GyrA, ParC) of 20 quinolone resistant E. coli isolates were identified by PCR and direct DNA sequencing. AcrA level was measured by real time PCR. GAPDH of E.coli was used as endogenous control. The expression of acrA was confirmed through northern hybridization method, the results obtained by real time PCR were compared.
Results
1) Topoisomerase mutations were found in all quinolone resistant E. coli strains. 2) AcrA expression in fluoroquinolone-resistant E. coli was quantified by using real time PCR. There was no relationship between the ratio of acrA expression to GAPDH and MIC of ciprofloxacin. 3) With Northern hybridization, we compared the band of acrA to that of GAPDH in compactness and area. No difference in the expression according to MIC could be found. 4) The results of AcrA/GAPDH were significantly correlated between the real-time PCR and northern blot (P<0.05, correlation coefficiency 0.98).
Conclusion
In this study, no relationship between overexpression of AcrA gene and high level fluoroquinolone resistance. Therefore, we assume that mechanism other than AcrAB efflux pump is involved in and contribute to high-level fluoroquinolone resistance. However, the degree of efflux pump expression could be confirmed with real time PCR using acrA mRNA. Therefore, real time PCR could be used in the molecular biologic study on the mechanism of resistance to antibiotics.
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Collections - 1. Basic Science > Department of Biochemistry and Molecular Biology > 1. Journal Articles
- 4. Research institute > Healthcare Readiness Institute for Unified Korea > 1. Journal Articles
- 2. Clinical Science > Department of Infectious Diseases > 1. Journal Articles
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