J Bacteriol Virol.  2019 Sep;49(3):95-114. 10.4167/jbv.2019.49.3.95.

Diversity of Genetic Environment of bla(CTX-M) Genes and Antimicrobial Susceptibility in Extended-spectrum β-lactamase producing Escherichia coli Isolated in Korea

• Yang YY
• Suh MH

• KMID: 2468030
• DOI: http://doi.org/10.4167/jbv.2019.49.3.95

Abstract

Increasing resistance due to the production of extended-spectrum β-lactamase (ESBL) in Escherichia coli is a major problem to public health and CTX-M enzymes have become the most prevalent ESBL worldwide. In this study, resistance profiles of E. coli isolated in Korea and the genetic environments of bla(CTX-M) genes were analyzed by PCR and direct sequencing to clarify the mechanisms of spread of CTX-M. Resistance rates of CTX-M-producing E. coli, including β-lactams, fluoroquinolones and aminoglycosides, were significantly higher than that of CTX-M-non-producers (p<0.01). Of 41 tested, 39 (95.1%) isolates of CTX-M-producing E. coli showed resistance transfer by conjugation. All the transconjugants harboured large plasmids of 118~172 megadalton. Insertion sequence ISEcp1B was detected in the upstream of the bla(CTX-M) in 38 (92.7%) isolates with bla(CTX-M). ISEcp1B was disrupted by IS26 in 16 (39.0%) isolates with bla(CTX-M). ISEcp1B carried −35 and −10 promoter components between right inverted repeat (IRR) and the start codon of bla(CTX-M). orf477 or IS903D was observed in the downstream of the bla(CTX-M) in all the isolates with bla(CTX-M-3/15/55) or with bla(CTX-M-14/27), respectively. Sequence similar to IRR of ISEcp1B was located downstream of orf477. Target duplication sequences were detected both upstream of IRL and downstream of IRR. These results showed the involvement of ISEcp1B in the mobilization of the resistance genes. In conclusion, the surrounding DNAs of bla(CTX-M) genes were very diverse, and the spread and the expression of CTX-M may be deeply related with ISEcp1B. These informations will provide important knowledge to control the increase in CTX-M-ESBLs.

Keyword

ESBL; CTX-M; ISEcp1B; IS26; Escherichia coli

MeSH Terms

Aminoglycosides
Codon, Initiator
DNA
Escherichia coli*
Escherichia*
Fluoroquinolones
Korea*
Plasmids
Polymerase Chain Reaction
Public Health
Aminoglycosides
Codon, Initiator
DNA
Fluoroquinolones

Figure

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  Figure 1. Resistance to increasing numbers of different antibiotics in CTX-M-producing (░) and CTX-M-non-producing (■) isolates.

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  Figure 2. Schematic representation of the surrounding DNA sequences of the blaCTx-m-3 genes of four multidrug resistant Escherichia coli isolates. Row1, JE120: 2, JE125: 3, JE64: 4, JE62. TD, Target duplication. IRL and IRR, left and right inverted repeat sequences for ISEcp1B or IS26. tnpA, transposase gene. ATG, start codon. TAA, stop codon. P, promotor.

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  Figure 3. Nucleotide sequence of DNA region of CTX-M-3-producing Escherichia coli JE1 20. The deduced amino acid sequence is indicated in single-letter code below the nucleotide sequence. Horizontal arrows, start codons. Asterisks, stop codons. TD, target duplication. IRL and IRR, left and right inverted repeat sequences, respectively. RBS, ribosomal binding site. The -35 and -10 signals, the promoter sequences of the blacTx-M-3. tnpA, transposase gene.

• 

  Figure 4. Schematic representation of the surrounding DNA sequences of the blacTx-M-15 and blacTx-M-55 genes of five multidrug resistant Escherichia coli isolates. Row1, JE36: 2, JE56 and JE11: 3, JE77: 4, JE28. TD, Target duplication. IRL and IRR, left and right inverted repeat sequences for IS26 or ISEcp1B. tnpA, transposase gene. ATG, start codon. TAA, stop codon P, promotor. a. TTCCT in JE11 (CTX-M-55). b. 45 bp in JE11 (CTX-M-55).

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  Figure 5. Schematic representation of the surrounding DNA sequences of the blacTx-M-14 genes of four multidrug resistant Escherichia coli isolates. Row1, JE45: 2, JE121: 3, JE123: 4, JE124. TD, Target duplication. IRL and IRR, left and right inverted repeat sequences for ISEcp/Bor IS26. tnpA, transposase gene. ATG and GTG, start codon. TAA, stop codon. P, promotor.

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  Figure 6. Schematic representation of the surrounding DNA sequences of the blactx-m-27 genes of three multidrug resistant E scherichia coli isolates. Row1, JE22: 2, JE78: 3, JE80. TD, Target duplication. IRL and IRR, left and right inverted repeat sequences for ISEcp/Bor IS26. tnpA, transposase gene. iron, iron transport gene. ATG and GTG, start codon. TAA, stop codon. P, promotor.

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