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

Abstract

Abstract 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-β-Lactamase-1 gene (bla NDM−1) in the clinical isolate (Klebsiella pneumoniae TH-P12158), transformant strains Escherichia coli BL21 (DE3)-bla NDM−1, and Escherichia coli DH5α- bla NDM−1 when exposed to imipenem. Methods β-Lactamase genes (bla SHV, bla TEM−1, bla CTX−M−9, bla IMP, bla NDM−1, bla KPC, bla OXA, bla GES, and bla 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 NDM−1 was transformed into E.coli BL21 (DE3) and E.coli DH5α by electroporation. The resistance phenotype and higher bla NDM−1 expression in K.pneumoniae TH-P12158, transformant E.coli BL21 (DE3)-bla NDM−1, and E.coli DH5α-bla 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 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 NDM−1 expression was deteriorated, while the MIC and MBC values remained relatively stable. These results demonstrated that low doses of imipenem (˂MIC) could press bla NDM−1 positive strains producing stable drug resistance memory and altered bla NDM−1 expression. Conclusions Low doses of imipenem could press bla NDM−1 positive strains producing sustained resistance memory and altered bla 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
• β-lactamase
• bla NDM−1
• Imipenem
• Drug resistance