Evolution of carbapenem resistance in klebsiella pneumoniae and escherichia coli carrying blaNDM−1 gene: imipenem exposure results in sustained resistance memory of strains in vitro

• Published in: Annals of clinical microbiology and antimicrobials 2023-06, Vol.22 (1), p.1-46, Article 46
• Main Authors: Zhao, Qiong, Sha, Longhua, Wu, Zhaomeng, Meng, Lixue, Yang, Feixiang, Wu, Lingling, Yu, Chunfang, Zhang, Hua, Yu, Jingdan, Jin, Zhixiong
• Format: Article
• Language: English
• Subjects: Analysis; Antibiotics; Antimicrobial agents; Bacteria; Bacterial infections; Beta lactamases; Carbapenems; Control; Cross infection; Drug resistance; Drug resistance in microorganisms; E coli; Electroporation; Enzymes; Escherichia coli; Evolution; Exposure; Gene expression; Gene mutations; Genes; Gram-negative bacteria; Health aspects; Identification and classification; Imipenem; Klebsiella; Laboratories; Metallo-β-lactamase; Metallography; Minimum inhibitory concentration; Nosocomial infections; Phenotypes; Plasmids; Prevention; Risk factors; Sodium; Strains (organisms); Subcultures
• ISSN: 1476-0711; 1476-0711
• DOI: 10.1186/s12941-023-00598-8

Background Antibiotics exert an outstanding selective pressure on bacteria, forcing their chromosomal gene mutations and drug resistance genes to spread. The objective of this study is to evaluate the expression of the New Delhi Metallo-[beta]-Lactamase-1 gene (bla.sub.NDM-1) in the clinical isolate (Klebsiella pneumoniae TH-P12158), transformant strains Escherichia coli BL21 (DE3)-bla.sub.NDM-1, and Escherichia coli DH5[alpha]- bla.sub.NDM-1 when exposed to imipenem. Methods [beta]-Lactamase genes (bla.sub.SHV, bla.sub.TEM-1, bla.sub.CTX-M-9, bla.sub.IMP, bla.sub.NDM-1, bla.sub.KPC, bla.sub.OXA, bla.sub.GES, and bla.sub.DHA) from randomly selected carbapenems-sensitive K.pneumoniae (n = 20) and E.coli (n = 20) strains were amplified by PCR. The recombinant plasmid of pET-28a harboring bla.sub.NDM-1 was transformed into E.coli BL21 (DE3) and E.coli DH5[alpha] by electroporation. The resistance phenotype and higher bla.sub.NDM-1 expression in K.pneumoniae TH-P12158, transformant E.coli BL21 (DE3)-bla.sub.NDM-1, and E.coli DH5[alpha]-bla.sub.NDM-1 were observed when exposed to imipenem with grade increasing, decreasing, and canceling doses, respectively. Results After being exposed to different doses of imipenem, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of antimicrobial drugs and bla.sub.NDM-1 expression of strains increased, which was positively correlated with doses of imipenem. On the contrary, with the decrease or cancellation of imipenem doses, the bla.sub.NDM-1 expression was deteriorated, while the MIC and MBC values remained relatively stable. These results demonstrated that low doses of imipenem (ËMIC) could press bla.sub.NDM-1 positive strains producing stable drug resistance memory and altered bla.sub.NDM-1 expression. Conclusions Low doses of imipenem could press bla.sub.NDM-1 positive strains producing sustained resistance memory and altered bla.sub.NDM-1 expression. In particular, the positive correlation between the resistance genes expression and antibiotics exposure shows promising guiding significance for clinical medication. Keywords: Klebsiella pneumoniae, Escherichia coli, [beta]-lactamase, bla.sub.NDM-1, Imipenem, Drug resistance