Emergence and characterization of IncFII/IncR plasmids with multiple 5,692 bp- blaKPC−2-bearing tandem repeats in ceftazidime/avibactam non-susceptible Klebsiella pneumoniae strains
Ceftazidime/avibactam (CAZ/AVI) is widely recognized as an effective treatment for infections caused by KPC-producing Klebsiella pneumoniae (KPC-Kp). However, the prevalence of CAZ/AVI resistance among KPC-Kp isolates has increased rapidly in recent years. In this study, high-level carbapenem resist...
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Frontiers Media S.A.
2025-04-01
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| author | Hongmao Liu Hongmao Liu Mei Zhu Junwan Lu Junwan Lu Shan Wu Rujian Ye Wei Pan Yirong Li Qiyu Bao Qiyu Bao Dawei Huang |
| author_facet | Hongmao Liu Hongmao Liu Mei Zhu Junwan Lu Junwan Lu Shan Wu Rujian Ye Wei Pan Yirong Li Qiyu Bao Qiyu Bao Dawei Huang |
| author_sort | Hongmao Liu |
| collection | DOAJ |
| description | Ceftazidime/avibactam (CAZ/AVI) is widely recognized as an effective treatment for infections caused by KPC-producing Klebsiella pneumoniae (KPC-Kp). However, the prevalence of CAZ/AVI resistance among KPC-Kp isolates has increased rapidly in recent years. In this study, high-level carbapenem resistance and enhanced CAZ/AVI resistance were observed in two hypervirulent carbapenem-resistant K. pneumoniae isolates, KP1878 and KP3034, following prolonged carbapenem use. Virulence phenotypes were confirmed using the string test and a Galleria mellonella larvae infection model. Real-time quantitative PCR revealed that the relative expression of blaKPC−2 in KP1878 and KP3034 was 2.4-fold and 11.6-fold higher, respectively, than that in the CAZ/AVI-susceptible KPC-Kp strain KP1880. Whole-genome sequencing showed that the blaKPC−2 gene resided within an identical 5,692-bp ΔklcA-korC-ΔISKpn6-blaKPC−2-ISKpn8-ΔtnpR-IS26 tandem repeat, which was replicated twice and four times in plasmids pKPC1878 and pKPC3034, respectively. Compared with KP1880, the β-lactamase hydrolysis activities of crude cell lysates derived from KP1878 and KP3034 were significantly higher in their ability to hydrolyze meropenem, ceftazidime, and nitrocefin. S1-nuclease-digested pulsed-field gel electrophoresis, along with Southern blot and restriction fragment length polymorphism fingerprinting, identified plasmid profiles but revealed one or more 5.6-kilobase variations in the regions hybridized with the KPC-specific probe. Further comparative genomic analysis suggested that a potential homologous recombination event occurred between the blaKPC−2-carrying plasmid and the pLVPK-like virulence plasmid of KP3034, leading to the generation of a cointegrated plasmid that combined both virulence and CAZ/AVI resistance. |
| format | Article |
| id | doaj-art-b7e99c1b293b48c8a28adf00a2f60a79 |
| institution | DOAJ |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj-art-b7e99c1b293b48c8a28adf00a2f60a792025-08-20T03:03:03ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-04-011610.3389/fmicb.2025.15346311534631Emergence and characterization of IncFII/IncR plasmids with multiple 5,692 bp- blaKPC−2-bearing tandem repeats in ceftazidime/avibactam non-susceptible Klebsiella pneumoniae strainsHongmao Liu0Hongmao Liu1Mei Zhu2Junwan Lu3Junwan Lu4Shan Wu5Rujian Ye6Wei Pan7Yirong Li8Qiyu Bao9Qiyu Bao10Dawei Huang11Department of Laboratory Medicine, Zhongnan Hospital of Wuhan University, Wuhan, ChinaSchool of Laboratory Medicine and Life Sciences, Institue of Biomedical Informatics, Wenzhou Medical University, Wenzhou, ChinaDepartment of Clinical Laboratory, Zhejiang Hospital, Hangzhou, ChinaSchool of Laboratory Medicine and Life Sciences, Institue of Biomedical Informatics, Wenzhou Medical University, Wenzhou, ChinaMedical Molecular Biology Laboratory, School of Medicine, Jinhua University of Vocational Technology, Jinhua, ChinaDepartment of Clinical Laboratory, The People's Hospital of Yuhuan, Taizhou, ChinaDepartment of Clinical Laboratory, The People's Hospital of Yuhuan, Taizhou, ChinaDepartment of Clinical Laboratory, The People's Hospital of Yuhuan, Taizhou, ChinaDepartment of Laboratory Medicine, Zhongnan Hospital of Wuhan University, Wuhan, ChinaSchool of Laboratory Medicine and Life Sciences, Institue of Biomedical Informatics, Wenzhou Medical University, Wenzhou, ChinaMedical Molecular Biology Laboratory, School of Medicine, Jinhua University of Vocational Technology, Jinhua, ChinaDepartment of Clinical Laboratory, The People's Hospital of Yuhuan, Taizhou, ChinaCeftazidime/avibactam (CAZ/AVI) is widely recognized as an effective treatment for infections caused by KPC-producing Klebsiella pneumoniae (KPC-Kp). However, the prevalence of CAZ/AVI resistance among KPC-Kp isolates has increased rapidly in recent years. In this study, high-level carbapenem resistance and enhanced CAZ/AVI resistance were observed in two hypervirulent carbapenem-resistant K. pneumoniae isolates, KP1878 and KP3034, following prolonged carbapenem use. Virulence phenotypes were confirmed using the string test and a Galleria mellonella larvae infection model. Real-time quantitative PCR revealed that the relative expression of blaKPC−2 in KP1878 and KP3034 was 2.4-fold and 11.6-fold higher, respectively, than that in the CAZ/AVI-susceptible KPC-Kp strain KP1880. Whole-genome sequencing showed that the blaKPC−2 gene resided within an identical 5,692-bp ΔklcA-korC-ΔISKpn6-blaKPC−2-ISKpn8-ΔtnpR-IS26 tandem repeat, which was replicated twice and four times in plasmids pKPC1878 and pKPC3034, respectively. Compared with KP1880, the β-lactamase hydrolysis activities of crude cell lysates derived from KP1878 and KP3034 were significantly higher in their ability to hydrolyze meropenem, ceftazidime, and nitrocefin. S1-nuclease-digested pulsed-field gel electrophoresis, along with Southern blot and restriction fragment length polymorphism fingerprinting, identified plasmid profiles but revealed one or more 5.6-kilobase variations in the regions hybridized with the KPC-specific probe. Further comparative genomic analysis suggested that a potential homologous recombination event occurred between the blaKPC−2-carrying plasmid and the pLVPK-like virulence plasmid of KP3034, leading to the generation of a cointegrated plasmid that combined both virulence and CAZ/AVI resistance.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1534631/fullKlebsiella pneumoniaeCeftazidime/avibactamblaKPC-2hypervirulent5,692 bp-tandem repeat |
| spellingShingle | Hongmao Liu Hongmao Liu Mei Zhu Junwan Lu Junwan Lu Shan Wu Rujian Ye Wei Pan Yirong Li Qiyu Bao Qiyu Bao Dawei Huang Emergence and characterization of IncFII/IncR plasmids with multiple 5,692 bp- blaKPC−2-bearing tandem repeats in ceftazidime/avibactam non-susceptible Klebsiella pneumoniae strains Frontiers in Microbiology Klebsiella pneumoniae Ceftazidime/avibactam blaKPC-2 hypervirulent 5,692 bp-tandem repeat |
| title | Emergence and characterization of IncFII/IncR plasmids with multiple 5,692 bp- blaKPC−2-bearing tandem repeats in ceftazidime/avibactam non-susceptible Klebsiella pneumoniae strains |
| title_full | Emergence and characterization of IncFII/IncR plasmids with multiple 5,692 bp- blaKPC−2-bearing tandem repeats in ceftazidime/avibactam non-susceptible Klebsiella pneumoniae strains |
| title_fullStr | Emergence and characterization of IncFII/IncR plasmids with multiple 5,692 bp- blaKPC−2-bearing tandem repeats in ceftazidime/avibactam non-susceptible Klebsiella pneumoniae strains |
| title_full_unstemmed | Emergence and characterization of IncFII/IncR plasmids with multiple 5,692 bp- blaKPC−2-bearing tandem repeats in ceftazidime/avibactam non-susceptible Klebsiella pneumoniae strains |
| title_short | Emergence and characterization of IncFII/IncR plasmids with multiple 5,692 bp- blaKPC−2-bearing tandem repeats in ceftazidime/avibactam non-susceptible Klebsiella pneumoniae strains |
| title_sort | emergence and characterization of incfii incr plasmids with multiple 5 692 bp blakpc 2 bearing tandem repeats in ceftazidime avibactam non susceptible klebsiella pneumoniae strains |
| topic | Klebsiella pneumoniae Ceftazidime/avibactam blaKPC-2 hypervirulent 5,692 bp-tandem repeat |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1534631/full |
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