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A Newly Discovered Drug Resistance Gene rfaF In Helicobacter pylori.
Infect Drug Resist 2019; 12:3507-3514ID

Abstract

Background

The purpose of this study was to understand the function of rfaF gene in Helicobacter pylori antibiotic resistance.

Methods

The gene homologous recombination method was used for knockout and complementation of H. pylori rfaF gene. Various constructed strains were analysed for drug sensitivity to amoxicillin (AMO), tetracycline (TET), clarithromycin (CLA), metronidazole (MET), levofloxacin (LEV), and chloramphenicol (CHL) by agar plate dilution method. Drug sensitivity was further confirmed using a growth inhibition curve. Ethidium bromide (EB) accumulation experiments were performed to assess cell membrane permeability. PCR and sequence analysis were used to detect the rfaF gene.

Results

The minimum inhibitory concentrations (MIC) of TET, CHL, AMO, and CLA in 11,637 rfaF knockout strain (ΔrfaF strain) were 4, 4, 2, and 2 times higher than those in 11,637 wild type (WT) strain, respectively. A multidrug-resistant (MDR) ΔrfaF strain also displayed the same trend; however, the degrees of increase were relatively small. Growth inhibition experiments indicated that the growth of the 11,637 ΔrfaF strain was higher with antibiotics at the MIC of the 11,637 WT strain than that of 11,637 rfaF-complemented strain (ΔrfaF/rfaF strain), whereas the 11,637 WT strain did not exhibit any growth. The 11,637 ΔrfaF strain was significantly reduced compared with the cumulative EB fluorescence intensity of the 11,637 WT and of 11,637ΔrfaF/rfaF strain, and the same trend appeared in the MDR strain. Among the 10 clinical strains, 9 clinical strains were found to have mutations in the conserved sequence of rfaF amino acids.

Conclusion

We found a new drug resistance gene, rfaF, in H. pylori, which changes the permeability of cell membrane to confer cross-resistance to AMO, TET, CLA, and CHL and is involved in clinical strain drug resistance. It can be used as a drug target.

Authors+Show Affiliations

Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China. Fujian Key Laboratory of Tumor Microbiology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China.Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China. Fujian Key Laboratory of Tumor Microbiology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China.Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China. Fujian Key Laboratory of Tumor Microbiology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China.Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China. Fujian Key Laboratory of Tumor Microbiology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China.Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China. Fujian Key Laboratory of Tumor Microbiology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, People's Republic of China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31814739

Citation

Lin, Jiansheng, et al. "A Newly Discovered Drug Resistance Gene rfaF in Helicobacter Pylori." Infection and Drug Resistance, vol. 12, 2019, pp. 3507-3514.
Lin J, Zhang X, Wen Y, et al. A Newly Discovered Drug Resistance Gene rfaF In Helicobacter pylori. Infect Drug Resist. 2019;12:3507-3514.
Lin, J., Zhang, X., Wen, Y., Chen, H., & She, F. (2019). A Newly Discovered Drug Resistance Gene rfaF In Helicobacter pylori. Infection and Drug Resistance, 12, pp. 3507-3514. doi:10.2147/IDR.S231152.
Lin J, et al. A Newly Discovered Drug Resistance Gene rfaF in Helicobacter Pylori. Infect Drug Resist. 2019;12:3507-3514. PubMed PMID: 31814739.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A Newly Discovered Drug Resistance Gene rfaF In Helicobacter pylori. AU - Lin,Jiansheng, AU - Zhang,Xiaoyan, AU - Wen,Yancheng, AU - Chen,Hao, AU - She,Feifei, Y1 - 2019/11/12/ PY - 2019/09/16/received PY - 2019/10/30/accepted PY - 2019/12/10/entrez PY - 2019/12/10/pubmed PY - 2019/12/10/medline KW - amoxicillin KW - clarithromycin KW - resistance KW - rfaF KW - tetracycline SP - 3507 EP - 3514 JF - Infection and drug resistance JO - Infect Drug Resist VL - 12 N2 - Background: The purpose of this study was to understand the function of rfaF gene in Helicobacter pylori antibiotic resistance. Methods: The gene homologous recombination method was used for knockout and complementation of H. pylori rfaF gene. Various constructed strains were analysed for drug sensitivity to amoxicillin (AMO), tetracycline (TET), clarithromycin (CLA), metronidazole (MET), levofloxacin (LEV), and chloramphenicol (CHL) by agar plate dilution method. Drug sensitivity was further confirmed using a growth inhibition curve. Ethidium bromide (EB) accumulation experiments were performed to assess cell membrane permeability. PCR and sequence analysis were used to detect the rfaF gene. Results: The minimum inhibitory concentrations (MIC) of TET, CHL, AMO, and CLA in 11,637 rfaF knockout strain (ΔrfaF strain) were 4, 4, 2, and 2 times higher than those in 11,637 wild type (WT) strain, respectively. A multidrug-resistant (MDR) ΔrfaF strain also displayed the same trend; however, the degrees of increase were relatively small. Growth inhibition experiments indicated that the growth of the 11,637 ΔrfaF strain was higher with antibiotics at the MIC of the 11,637 WT strain than that of 11,637 rfaF-complemented strain (ΔrfaF/rfaF strain), whereas the 11,637 WT strain did not exhibit any growth. The 11,637 ΔrfaF strain was significantly reduced compared with the cumulative EB fluorescence intensity of the 11,637 WT and of 11,637ΔrfaF/rfaF strain, and the same trend appeared in the MDR strain. Among the 10 clinical strains, 9 clinical strains were found to have mutations in the conserved sequence of rfaF amino acids. Conclusion: We found a new drug resistance gene, rfaF, in H. pylori, which changes the permeability of cell membrane to confer cross-resistance to AMO, TET, CLA, and CHL and is involved in clinical strain drug resistance. It can be used as a drug target. SN - 1178-6973 UR - https://www.unboundmedicine.com/medline/citation/31814739/A_Newly_Discovered_Drug_Resistance_Gene_rfaF_In_Helicobacter_pylori L2 - https://dx.doi.org/10.2147/IDR.S231152 DB - PRIME DP - Unbound Medicine ER -