Characterization of a mcr-1 and CRISPR-Cas System Co-harboring Plasmid in a Carbapenemase-Producing High-Risk ST11 Klebsiella pneumoniae Strain
We set out to study the prevalence of the gene in carbapenemase-producing (CPKP) strains, and to determine whether its presence is associated with a fitness cost. A total of 234 clinical CPKP isolates were collected from a tertiary medical center in Taiwan from January 2018 to January 2019. The and...
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| Published in | Frontiers in microbiology Vol. 12; p. 762947 |
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| Main Authors | , , , , , , , |
| Format | Journal Article |
| Language | English |
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27.10.2021
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| Abstract | We set out to study the prevalence of the
gene in carbapenemase-producing
(CPKP) strains, and to determine whether its presence is associated with a fitness cost. A total of 234 clinical CPKP isolates were collected from a tertiary medical center in Taiwan from January 2018 to January 2019. The
and carbapenemase genes were detected by polymerase chain reaction (PCR) followed by Sanger sequencing. The
-positive carbapenemase-producing strain was characterized by whole genome sequencing, a plasmid stability test and a conjugation assay.
growth rate and an
virulence test were compared between the parental
-positive strain and its
plasmid-cured strain. We identified only one
positive strain (KP2509), co-harboring
and
, among 234 (1/234, 0.43%) CPKP strains. KP2509 and its
transconjugant showed moderate colistin resistance (MIC = 8 mg/L). The
is located on a large conjugative plasmid (317 kb), pKP2509-MCR, with three replicons, IncHI, IncFIB, and IncN. Interestingly, a complete Type IV-A3 CRISPR-Cas system was identified in pKP2509-MCR. Plasmid pKP2509-MCR was highly stable in KP2509 after 270 generation of passage, and the pKP2509-MCR cured strain PC-KP2509 showed similar growth rate and
virulence in comparison to KP2509. The prevalence of
in CPKP strains remains low in our center. Notably, we identified a large plasmid with multiple replicons containing both the
and the Type IV-3A CRISPR-Cas genes. The further spread of this highly stable plasmid raises concern that it may promote the increase of
prevalence in CPKP. |
|---|---|
| AbstractList | We set out to study the prevalence of the
gene in carbapenemase-producing
(CPKP) strains, and to determine whether its presence is associated with a fitness cost. A total of 234 clinical CPKP isolates were collected from a tertiary medical center in Taiwan from January 2018 to January 2019. The
and carbapenemase genes were detected by polymerase chain reaction (PCR) followed by Sanger sequencing. The
-positive carbapenemase-producing strain was characterized by whole genome sequencing, a plasmid stability test and a conjugation assay.
growth rate and an
virulence test were compared between the parental
-positive strain and its
plasmid-cured strain. We identified only one
positive strain (KP2509), co-harboring
and
, among 234 (1/234, 0.43%) CPKP strains. KP2509 and its
transconjugant showed moderate colistin resistance (MIC = 8 mg/L). The
is located on a large conjugative plasmid (317 kb), pKP2509-MCR, with three replicons, IncHI, IncFIB, and IncN. Interestingly, a complete Type IV-A3 CRISPR-Cas system was identified in pKP2509-MCR. Plasmid pKP2509-MCR was highly stable in KP2509 after 270 generation of passage, and the pKP2509-MCR cured strain PC-KP2509 showed similar growth rate and
virulence in comparison to KP2509. The prevalence of
in CPKP strains remains low in our center. Notably, we identified a large plasmid with multiple replicons containing both the
and the Type IV-3A CRISPR-Cas genes. The further spread of this highly stable plasmid raises concern that it may promote the increase of
prevalence in CPKP. We set out to study the prevalence of the mcr-1 gene in carbapenemase-producing Klebsiella pneumoniae (CPKP) strains, and to determine whether its presence is associated with a fitness cost. A total of 234 clinical CPKP isolates were collected from a tertiary medical center in Taiwan from January 2018 to January 2019. The mcr-1 and carbapenemase genes were detected by polymerase chain reaction (PCR) followed by Sanger sequencing. The mcr-1 -positive carbapenemase-producing strain was characterized by whole genome sequencing, a plasmid stability test and a conjugation assay. In vitro growth rate and an in vivo virulence test were compared between the parental mcr-1 -positive strain and its mcr-1 plasmid-cured strain. We identified only one mcr-1 positive strain (KP2509), co-harboring bla KPC– 2 and bla OXA– 48 , among 234 (1/234, 0.43%) CPKP strains. KP2509 and its Escherichia coli mcr-1 transconjugant showed moderate colistin resistance (MIC = 8 mg/L). The mcr-1 is located on a large conjugative plasmid (317 kb), pKP2509-MCR, with three replicons, IncHI, IncFIB, and IncN. Interestingly, a complete Type IV-A3 CRISPR-Cas system was identified in pKP2509-MCR. Plasmid pKP2509-MCR was highly stable in KP2509 after 270 generation of passage, and the pKP2509-MCR cured strain PC-KP2509 showed similar growth rate and in vivo virulence in comparison to KP2509. The prevalence of mcr-1 in CPKP strains remains low in our center. Notably, we identified a large plasmid with multiple replicons containing both the mcr-1 and the Type IV-3A CRISPR-Cas genes. The further spread of this highly stable plasmid raises concern that it may promote the increase of mcr-1 prevalence in CPKP. We set out to study the prevalence of the mcr-1 gene in carbapenemase-producing Klebsiella pneumoniae (CPKP) strains, and to determine whether its presence is associated with a fitness cost. A total of 234 clinical CPKP isolates were collected from a tertiary medical center in Taiwan from January 2018 to January 2019. The mcr-1 and carbapenemase genes were detected by polymerase chain reaction (PCR) followed by Sanger sequencing. The mcr-1-positive carbapenemase-producing strain was characterized by whole genome sequencing, a plasmid stability test and a conjugation assay. In vitro growth rate and an in vivo virulence test were compared between the parental mcr-1-positive strain and its mcr-1 plasmid-cured strain. We identified only one mcr-1 positive strain (KP2509), co-harboring blaKPC–2 and blaOXA–48, among 234 (1/234, 0.43%) CPKP strains. KP2509 and its Escherichia coli mcr-1 transconjugant showed moderate colistin resistance (MIC = 8 mg/L). The mcr-1 is located on a large conjugative plasmid (317 kb), pKP2509-MCR, with three replicons, IncHI, IncFIB, and IncN. Interestingly, a complete Type IV-A3 CRISPR-Cas system was identified in pKP2509-MCR. Plasmid pKP2509-MCR was highly stable in KP2509 after 270 generation of passage, and the pKP2509-MCR cured strain PC-KP2509 showed similar growth rate and in vivo virulence in comparison to KP2509. The prevalence of mcr-1 in CPKP strains remains low in our center. Notably, we identified a large plasmid with multiple replicons containing both the mcr-1 and the Type IV-3A CRISPR-Cas genes. The further spread of this highly stable plasmid raises concern that it may promote the increase of mcr-1 prevalence in CPKP. We set out to study the prevalence of the mcr-1 gene in carbapenemase-producing Klebsiella pneumoniae (CPKP) strains, and to determine whether its presence is associated with a fitness cost. A total of 234 clinical CPKP isolates were collected from a tertiary medical center in Taiwan from January 2018 to January 2019. The mcr-1 and carbapenemase genes were detected by polymerase chain reaction (PCR) followed by Sanger sequencing. The mcr-1-positive carbapenemase-producing strain was characterized by whole genome sequencing, a plasmid stability test and a conjugation assay. In vitro growth rate and an in vivo virulence test were compared between the parental mcr-1-positive strain and its mcr-1 plasmid-cured strain. We identified only one mcr-1 positive strain (KP2509), co-harboring bla KPC- 2 and bla OXA- 48, among 234 (1/234, 0.43%) CPKP strains. KP2509 and its Escherichia coli mcr-1 transconjugant showed moderate colistin resistance (MIC = 8 mg/L). The mcr-1 is located on a large conjugative plasmid (317 kb), pKP2509-MCR, with three replicons, IncHI, IncFIB, and IncN. Interestingly, a complete Type IV-A3 CRISPR-Cas system was identified in pKP2509-MCR. Plasmid pKP2509-MCR was highly stable in KP2509 after 270 generation of passage, and the pKP2509-MCR cured strain PC-KP2509 showed similar growth rate and in vivo virulence in comparison to KP2509. The prevalence of mcr-1 in CPKP strains remains low in our center. Notably, we identified a large plasmid with multiple replicons containing both the mcr-1 and the Type IV-3A CRISPR-Cas genes. The further spread of this highly stable plasmid raises concern that it may promote the increase of mcr-1 prevalence in CPKP. |
| Author | Huang, Po-Han Kreiswirth, Barry N Chen, Liang Juan, Yu-Fan Yang, Tsuey-Ching Chou, Sheng-Hua Cheng, Yi-Hsiang Lin, Yi-Tsung |
| AuthorAffiliation | 5 Hackensack Meridian Health Center for Discovery and Innovation , Nutley, NJ , United States 1 Division of Infectious Diseases, Department of Medicine, Taipei Veterans General Hospital , Taipei , Taiwan 3 Department of Biotechnology and Laboratory Science in Medicine, National Yang Ming Chiao Tung University , Taipei , Taiwan 2 Institute of Emergency and Critical Care Medicine, National Yang Ming Chiao Tung University , Taipei , Taiwan 6 Department of Medical Sciences, Hackensack Meridian School of Medicine , Nutley, NJ , United States 4 Division of Microbiology, Department of Pathology and Laboratory Medicine, Taipei Veterans General Hospital , Taipei , Taiwan |
| AuthorAffiliation_xml | – name: 2 Institute of Emergency and Critical Care Medicine, National Yang Ming Chiao Tung University , Taipei , Taiwan – name: 4 Division of Microbiology, Department of Pathology and Laboratory Medicine, Taipei Veterans General Hospital , Taipei , Taiwan – name: 3 Department of Biotechnology and Laboratory Science in Medicine, National Yang Ming Chiao Tung University , Taipei , Taiwan – name: 5 Hackensack Meridian Health Center for Discovery and Innovation , Nutley, NJ , United States – name: 6 Department of Medical Sciences, Hackensack Meridian School of Medicine , Nutley, NJ , United States – name: 1 Division of Infectious Diseases, Department of Medicine, Taipei Veterans General Hospital , Taipei , Taiwan |
| Author_xml | – sequence: 1 givenname: Yi-Hsiang surname: Cheng fullname: Cheng, Yi-Hsiang organization: Institute of Emergency and Critical Care Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan – sequence: 2 givenname: Sheng-Hua surname: Chou fullname: Chou, Sheng-Hua organization: Institute of Emergency and Critical Care Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan – sequence: 3 givenname: Po-Han surname: Huang fullname: Huang, Po-Han organization: Division of Infectious Diseases, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan – sequence: 4 givenname: Tsuey-Ching surname: Yang fullname: Yang, Tsuey-Ching organization: Department of Biotechnology and Laboratory Science in Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan – sequence: 5 givenname: Yu-Fan surname: Juan fullname: Juan, Yu-Fan organization: Division of Microbiology, Department of Pathology and Laboratory Medicine, Taipei Veterans General Hospital, Taipei, Taiwan – sequence: 6 givenname: Barry N surname: Kreiswirth fullname: Kreiswirth, Barry N organization: Hackensack Meridian Health Center for Discovery and Innovation, Nutley, NJ, United States – sequence: 7 givenname: Yi-Tsung surname: Lin fullname: Lin, Yi-Tsung organization: Institute of Emergency and Critical Care Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan – sequence: 8 givenname: Liang surname: Chen fullname: Chen, Liang organization: Department of Medical Sciences, Hackensack Meridian School of Medicine, Nutley, NJ, United States |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/34777318$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_1016_j_molcel_2023_05_036 crossref_primary_10_1128_aac_01189_22 crossref_primary_10_1016_j_jgar_2023_04_009 |
| Cites_doi | 10.1093/nar/gkz1197 10.3389/fmicb.2020.01937 10.1093/cid/cix893 10.1016/S1473-3099(16)30527-8 10.1093/jac/dky164 10.1089/crispr.2019.0048 10.3389/fmicb.2019.02934 10.1089/mdr.2017.0400 10.1038/s41467-017-02149-0 10.1006/abio.1995.1220 10.1128/AAC.00317-17 10.1016/S1473-3099(16)30528-X 10.1093/jac/dkx491 10.1128/IAI.00127-11 10.1038/s41579-019-0299-x 10.1016/j.ijantimicag.2016.06.023 10.1093/jac/dky061 10.1016/j.diagmicrobio.2017.09.016 10.1038/nrmicro3569 10.1093/nar/gkab149 10.1016/j.cmi.2020.07.043 10.1016/s1369-5274(99)00005-3 10.2147/IDR.S292820 10.3201/eid2404.171787 10.1016/j.ijantimicag.2017.11.011 10.1128/AAC.01075-16 10.1093/gbe/evx192 10.1093/femsre/fux013 10.1128/mSphere.00551-19 10.1016/S1473-3099(15)00424-7 10.1128/CMR.00064-16 |
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| Copyright | Copyright © 2021 Cheng, Chou, Huang, Yang, Juan, Kreiswirth, Lin and Chen. Copyright © 2021 Cheng, Chou, Huang, Yang, Juan, Kreiswirth, Lin and Chen. 2021 Cheng, Chou, Huang, Yang, Juan, Kreiswirth, Lin and Chen |
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| Keywords | fitness cost carbapenemase colistin high-risk clone mcr-1 |
| Language | English |
| License | Copyright © 2021 Cheng, Chou, Huang, Yang, Juan, Kreiswirth, Lin and Chen. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Reviewed by: Jens Andre Hammerl, Bundesinstitut für Risikobewertung, Germany; Jian-Hua Liu, South China Agricultural University, China; Chengming Wang, Auburn University, United States These authors have contributed equally to this work Edited by: Miklos Fuzi, Semmelweis University, Hungary This article was submitted to Antimicrobials, Resistance and Chemotherapy, a section of the journal Frontiers in Microbiology |
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| Snippet | We set out to study the prevalence of the
gene in carbapenemase-producing
(CPKP) strains, and to determine whether its presence is associated with a fitness... We set out to study the prevalence of the mcr-1 gene in carbapenemase-producing Klebsiella pneumoniae (CPKP) strains, and to determine whether its presence is... We set out to study the prevalence of the mcr-1 gene in carbapenemase-producing Klebsiella pneumoniae (CPKP) strains, and to determine whether its presence is... |
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| SubjectTerms | carbapenemase colistin fitness cost high-risk clone mcr-1 Microbiology |
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| Title | Characterization of a mcr-1 and CRISPR-Cas System Co-harboring Plasmid in a Carbapenemase-Producing High-Risk ST11 Klebsiella pneumoniae Strain |
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