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Published on: March 24, 2023
Description of novel resistance islands harbouring blaCTX-M-2 in IncC type 2 plasmids
R Papa-Ezdra1, N F Cordeiro1, V Di Pilato2
1Departamento de Bacteriología y Virología, Instituto de Higiene, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
Objectives:
We sequenced two IncA/C plasmids harbouring blaCTX-M-2 in Klebsiella pneumoniae clinical isolates and compared their antibiotic resistance islands.
Methods:
Transconjugants were obtained from two clinical K. pneumoniae isolates harbouring blaCTX-M-2. Plasmid DNA from transconjugants underwent short-read whole-genome sequencing, reads were assembled, and gaps were closed by PCR and sequencing. Determination of plasmid replicons, antibiotic resistance genes, identification and characterisation of insertion sequence (IS) elements, and comparison with publicly available plasmid sequences were performed.
Results:
blaCTX-M-2 was located in a complex class 1 integron In35::ISCR1::blaCTX-M-2, inserted in two different transposons designated Tn7057 and Tn7058, that reside in the resistance islands of plasmids pUR-KP0923 and pUR-KP1025, respectively. The general modules of both transposons were In35::ISCR1::blaCTX-M-2-Tn1000-like-Tn2*-ISKpn11-12-13 variable module-ΔTn21. In Tn7057 there was ΔIS10R-catA2 associated with an additional ISKpn13. Both plasmids belonged to IncC type 2 and ST3. pUR-KP0923 was 167 138 bp in length and had a 37 926-bp resistance island at position 4 (RI-4). Plasmid pUR-KP1025 was 168 128 bp with a RI-4 of 36 222 bp.
Conclusion:
This report describes the molecular nature of two transposons (Tn7057 and Tn7058) harbouring blaCTX-M-2 that reside in IncC type 2 ST3 plasmids. These transposons mediate resistance to oxyimino-cephalosporins, gentamicin and, in the case of Tn7057, chloramphenicol. CTX-M-2 is an important extended-spectrum β-lactamase (ESBL) to South American epidemiology. It is remarkable that despite being only two plasmid sequences, the information revealed here could contribute to a better understanding of the resistance islands from IncC type 2 plasmids.
Insights
Two Klebsiella pneumoniae plasmids carrying blaCTX-M-2 were sequenced, revealing complex transposons Tn7057 and Tn7058. These IncC plasmids contribute to understanding antibiotic resistance islands.
Area of Science:
- Microbiology
- Genetics
Background:
- Extended-spectrum β-lactamases (ESBLs) like CTX-M-2 are significant in clinical settings.
- IncA/C plasmids are known vehicles for antibiotic resistance gene dissemination.
Purpose of the Study:
- To characterize the molecular structure of two IncA/C plasmids harbouring blaCTX-M-2 in Klebsiella pneumoniae.
- To compare the antibiotic resistance islands within these plasmids.
Main Methods:
- Whole-genome sequencing of transconjugant plasmids.
- Plasmid assembly, gap closure via PCR and sequencing.
- Identification of replicons, resistance genes, and insertion sequence elements.
Main Results:
- The blaCTX-M-2 gene was found within complex class 1 integrons inserted into two distinct transposons, Tn7057 and Tn7058.
- Both transposons shared common modules but had variable regions, with Tn7057 containing additional resistance genes.
- The plasmids, pUR-KP0923 and pUR-KP1025, were identified as IncC type 2 ST3, with large resistance islands (RI-4).
Conclusions:
- The study details the molecular architecture of two novel transposons, Tn7057 and Tn7058, carrying blaCTX-M-2 on IncC plasmids.
- These transposons confer resistance to multiple antibiotics, including oxyimino-cephalosporins and gentamicin.
- The findings enhance understanding of resistance island evolution within IncC type 2 plasmids, relevant to South American epidemiology.
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