Infection and Drug Resistance (Sep 2019)
The resistance mechanism of Escherichia coli induced by ampicillin in laboratory
Abstract
Mengchen Li,1 Qiaoli Liu,1 Yanli Teng,1 Liuyang Ou,1 Yuanlin Xi,1 Shuaiyin Chen,1,* Guangcai Duan1,2,* 1Department of Epidemiology, College of Public Health, Zhengzhou University, Zhengzhou, Henan, People’s Republic of China; 2Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, Xinxiang Medical College, Xinxiang, Henan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Shuaiyin Chen; Guangcai DuanDepartment of Epidemiology, College of Public Health, Zhengzhou University, No.100 Kexue Avenue, Zhengzhou, Henan 450001, People’s Republic of ChinaTel +86 1 352 340 8394Fax +86 6 778 1453Email [email protected]; [email protected]: Multi-drug-resistant Escherichia coli poses a great threat to human health, especially resistant to ampicillin (AMP), but the mechanism of drug resistance is not very clear.Purpose: To understand the mechanism of resistance of E. coli to beta-lactam antibiotics by inducing drug resistance of sensitive bacteria in laboratory.Methods: Clinical sensitive E. coli strain was induced into resistance strain by 1/2 minimum inhibitive concentration (MIC) induced trails of AMP. The drug resistance spectrum was measured by modified K-B susceptibility test. Whole-genome sequencing analysis was used to analyze primary sensitive strain, and resequencing was used to analyze induced strains. Protein tertiary structure encoded by the gene containing single nucleotide polymorphism (SNP) was analyzed by bioinformatics.Results: After 315 hrs induced, the MIC value of E. coli 15743 reached to 256 μg/mL, 64 times higher than that of the sensitive bacteria. During the induction process, the bacterial resistance process is divided into two stages. The rate of drug resistance occurs rapidly before reaching the critical concentration of 32 μg/mL, and then the resistance rate slows down. Sequencing of the genome of resistant strain showed that E. coli 15743 drug-resistant strain with the MIC values of 32 and 256 μg/mL contained four and eight non-synonymous SNPs, respectively. These non-synonymous SNPs were distributed in the genes of frdD, ftsI, acrB, OmpD, marR, VgrG, and envZ.Conclusion: These studies will improve our understanding of the molecular mechanism of AMP resistance of E. coli, and may provide the basis for prevention and control of multi-drug-resistant bacteria and generation of new antibiotics to treat E. coli infection.Keywords: Escherichia coli, ampicillin, drug resistance
