{"title":"意大利重症监护病房爆发的耐头孢他啶/阿维菌素 ST307 肺炎克雷伯菌中的新型 KPC-166","authors":"Aurora Piazza , Vittoria Mattioni Marchetti , Alessandra Bielli , Gherard Batisti Biffignandi , Francesca Piscopiello , Riccardo Giudici , Livia Tartaglione , Marco Merli , Chiara Vismara , Roberta Migliavacca","doi":"10.1016/j.jmii.2024.03.004","DOIUrl":null,"url":null,"abstract":"<div><h3>Introduction</h3><p>Aim of the study was the molecular characterization of 21 ceftazidime/avibactam resistant (CZA-R) <em>Klebsiella pneumoniae</em> strains, collected in the period October 2021–March 2022 from an Intensive Care COVID Unit in a Northern Italian Hospital.</p></div><div><h3>Methods</h3><p>After growth on selective/chromogenic culture media and susceptibility tests assessment, resistance genes content was ascertained for all the isolates by the HybriSpot 12 multiplexing, PCR and Whole-Genome Sequencing (WGS). Clonality was assessed by PFGE and MLST according to the Pasteur scheme. A SNPs-based phylogenetic tree was obtained comparing representative isolates and global genomes. The <em>bla</em>KPC gene horizontal transmission was evaluated by conjugation experiments. <em>bla</em>KPC-166 was cloned in a pCR2.1 vector and transformed in chemically competent TOP10 cells.</p></div><div><h3>Results</h3><p>Sixteen inpatients resulted positive for colonization and/or infection by KPC-producing <em>K. pneumoniae</em> (KPC-Kp) strains. The 21 CZA-R KPC-Kp isolates obtained showed MDR phenotype; susceptibility to meropenem was always retained. All the CZA-R KPC-Kp presented a novel <em>bla</em>KPC variant, named <em>bla</em>KPC-166, showing a single nucleotide substitution (T811C) compared to the <em>bla</em>KPC-94; but related to <em>bla</em>KPC-2.</p></div><div><h3>Two different pulsotypes were detected</h3><p>A in 18/21 and B in 1/21 cases, two strains from the same patient being untypable by PFGE. Interestingly, the outbreak was sustained by the high-risk clone ST307, although the ST22, ST6342, ST6418 and ST6811 have also been identified and associated to KPC-166. Worryingly, <em>bla</em>KPC-166 could be transferred horizontally and, after cloning, it conferred resistance to CZA.</p></div><div><h3>Discussion</h3><p>This novel variant confers CZA–resistance and carbapenems susceptibility restoration. As KPC-166 was found expressed by multiple Kp clones, greater efforts should be made to prevent the further dissemination of such strains in Italian clinical settings.</p></div>","PeriodicalId":56117,"journal":{"name":"Journal of Microbiology Immunology and Infection","volume":"57 3","pages":"Pages 457-469"},"PeriodicalIF":4.5000,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1684118224000677/pdfft?md5=2e736469fac2a72172d5b86673192c7d&pid=1-s2.0-S1684118224000677-main.pdf","citationCount":"0","resultStr":"{\"title\":\"A novel KPC-166 in ceftazidime/avibactam resistant ST307 Klebsiella pneumoniae causing an outbreak in intensive care COVID Unit, Italy\",\"authors\":\"Aurora Piazza , Vittoria Mattioni Marchetti , Alessandra Bielli , Gherard Batisti Biffignandi , Francesca Piscopiello , Riccardo Giudici , Livia Tartaglione , Marco Merli , Chiara Vismara , Roberta Migliavacca\",\"doi\":\"10.1016/j.jmii.2024.03.004\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Introduction</h3><p>Aim of the study was the molecular characterization of 21 ceftazidime/avibactam resistant (CZA-R) <em>Klebsiella pneumoniae</em> strains, collected in the period October 2021–March 2022 from an Intensive Care COVID Unit in a Northern Italian Hospital.</p></div><div><h3>Methods</h3><p>After growth on selective/chromogenic culture media and susceptibility tests assessment, resistance genes content was ascertained for all the isolates by the HybriSpot 12 multiplexing, PCR and Whole-Genome Sequencing (WGS). Clonality was assessed by PFGE and MLST according to the Pasteur scheme. A SNPs-based phylogenetic tree was obtained comparing representative isolates and global genomes. The <em>bla</em>KPC gene horizontal transmission was evaluated by conjugation experiments. <em>bla</em>KPC-166 was cloned in a pCR2.1 vector and transformed in chemically competent TOP10 cells.</p></div><div><h3>Results</h3><p>Sixteen inpatients resulted positive for colonization and/or infection by KPC-producing <em>K. pneumoniae</em> (KPC-Kp) strains. The 21 CZA-R KPC-Kp isolates obtained showed MDR phenotype; susceptibility to meropenem was always retained. All the CZA-R KPC-Kp presented a novel <em>bla</em>KPC variant, named <em>bla</em>KPC-166, showing a single nucleotide substitution (T811C) compared to the <em>bla</em>KPC-94; but related to <em>bla</em>KPC-2.</p></div><div><h3>Two different pulsotypes were detected</h3><p>A in 18/21 and B in 1/21 cases, two strains from the same patient being untypable by PFGE. Interestingly, the outbreak was sustained by the high-risk clone ST307, although the ST22, ST6342, ST6418 and ST6811 have also been identified and associated to KPC-166. Worryingly, <em>bla</em>KPC-166 could be transferred horizontally and, after cloning, it conferred resistance to CZA.</p></div><div><h3>Discussion</h3><p>This novel variant confers CZA–resistance and carbapenems susceptibility restoration. As KPC-166 was found expressed by multiple Kp clones, greater efforts should be made to prevent the further dissemination of such strains in Italian clinical settings.</p></div>\",\"PeriodicalId\":56117,\"journal\":{\"name\":\"Journal of Microbiology Immunology and Infection\",\"volume\":\"57 3\",\"pages\":\"Pages 457-469\"},\"PeriodicalIF\":4.5000,\"publicationDate\":\"2024-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S1684118224000677/pdfft?md5=2e736469fac2a72172d5b86673192c7d&pid=1-s2.0-S1684118224000677-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Microbiology Immunology and Infection\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1684118224000677\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"IMMUNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Microbiology Immunology and Infection","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1684118224000677","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
A novel KPC-166 in ceftazidime/avibactam resistant ST307 Klebsiella pneumoniae causing an outbreak in intensive care COVID Unit, Italy
Introduction
Aim of the study was the molecular characterization of 21 ceftazidime/avibactam resistant (CZA-R) Klebsiella pneumoniae strains, collected in the period October 2021–March 2022 from an Intensive Care COVID Unit in a Northern Italian Hospital.
Methods
After growth on selective/chromogenic culture media and susceptibility tests assessment, resistance genes content was ascertained for all the isolates by the HybriSpot 12 multiplexing, PCR and Whole-Genome Sequencing (WGS). Clonality was assessed by PFGE and MLST according to the Pasteur scheme. A SNPs-based phylogenetic tree was obtained comparing representative isolates and global genomes. The blaKPC gene horizontal transmission was evaluated by conjugation experiments. blaKPC-166 was cloned in a pCR2.1 vector and transformed in chemically competent TOP10 cells.
Results
Sixteen inpatients resulted positive for colonization and/or infection by KPC-producing K. pneumoniae (KPC-Kp) strains. The 21 CZA-R KPC-Kp isolates obtained showed MDR phenotype; susceptibility to meropenem was always retained. All the CZA-R KPC-Kp presented a novel blaKPC variant, named blaKPC-166, showing a single nucleotide substitution (T811C) compared to the blaKPC-94; but related to blaKPC-2.
Two different pulsotypes were detected
A in 18/21 and B in 1/21 cases, two strains from the same patient being untypable by PFGE. Interestingly, the outbreak was sustained by the high-risk clone ST307, although the ST22, ST6342, ST6418 and ST6811 have also been identified and associated to KPC-166. Worryingly, blaKPC-166 could be transferred horizontally and, after cloning, it conferred resistance to CZA.
Discussion
This novel variant confers CZA–resistance and carbapenems susceptibility restoration. As KPC-166 was found expressed by multiple Kp clones, greater efforts should be made to prevent the further dissemination of such strains in Italian clinical settings.
期刊介绍：
Journal of Microbiology Immunology and Infection is an open access journal, committed to disseminating information on the latest trends and advances in microbiology, immunology, infectious diseases and parasitology. Article types considered include perspectives, review articles, original articles, brief reports and correspondence.
With the aim of promoting effective and accurate scientific information, an expert panel of referees constitutes the backbone of the peer-review process in evaluating the quality and content of manuscripts submitted for publication.
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