{"title":"开发并验证用于快速检测肠杆菌中 blaCTX-M、blaOXA-1、blaCMY、blaNDM 和 PBP3 插入物的五重 PCR 法。","authors":"Yamuna Devi Bakthavatchalam , Fizaa Abdullah , Devishree Srinivasan , Sangeetha Nithiyanandam , Ayyanraj Neeravi , Poojah Shah , Nivedhana Subburaju , Subha Vajjiravelu Jaganathan , Rema Devi , Gita Nataraj , Binesh Lal Yesudason , Kamini Walia , Balaji Veeraraghavan","doi":"10.1016/j.ijmmb.2024.100710","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><p>There is a high diversity of beta-lactamases in gram negative pathogens, making them difficult to treat. In the presence of OXA-1 and ampC, PTZ is no longer clinically relevant when treating Enterobacterales expressing ESBLs. Further, MBL infections are often treated with the combination of ceftazidime/avibactam with aztreonam. . It has recently been reported that NDM-expressing <em>E. coli</em> isolates co-harboring PBP3 insert develops resistance to this triple combination.</p></div><div><h3>Methods</h3><p>A pentaplex PCR is developed and validated to simultaneously detect bla<sub>CTX-M</sub>, bla<sub>OXA-1</sub>, bla<sub>CMY</sub>, bla<sub>NDM</sub>, and the PBP3 insert in whole genome sequenced <em>E. coli</em> and <em>K. pneumoniae</em> isolates. In addition, the isolates chosen for pentaplex PCR evaluation were tested for their minimum inhibitory concentrations (MICs) against piperacillin/tazobactam, cefoperazone/sulbactam (C/S), ertapenem, imipenem, meropenem, ceftazidime/avibactam, aztreonam/avibactam, cefepime/taniborbactam, and cefiderocol.</p></div><div><h3>Results</h3><p>The developed pentaplex PCR showed 100 % reproducibility with the antimicrobial resistance profile generated from whole genome sequenced data. PTZ and C/S are not effective against ESBL and/or OXA-1 expressing <em>E. coli</em> and <em>K. pneumoniae</em> isolates and do not offer any activity against CMY co-producers. Further, the combined effect of CMY, NDM and PBP3 inserts impacts aztreonam/avibactam activity and reduces the susceptibility to 40 % in <em>E. coli</em> isolates. While, aztreonam/avibactam showed potent activity against NDM-expressing <em>K. pneumoniae</em> isolates. Importantly, cefepime/taniborbactam and cefiderocol showed limited activity against NDM-expressing <em>E. coli</em> and <em>K. pneumoniae</em> isolates.</p></div><div><h3>Conclusion</h3><p>The pentaplex PCR was effective in detecting four beta-lactamases (bla<sub>CTX-M</sub>, bla<sub>OXA-1</sub>, bla<sub>CMY</sub>, bla<sub>NDM</sub>) as well as PBP3 inserts. It is expected that using pentaplex PCR as a diagnostic test for resistance detection in clinical practice will improve patient outcomes by providing prompt and targeted treatment.</p></div>","PeriodicalId":13284,"journal":{"name":"Indian Journal of Medical Microbiology","volume":null,"pages":null},"PeriodicalIF":1.4000,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development and validation of a pentaplex PCR assay for rapid detection of blaCTX-M, blaOXA–1, blaCMY, blaNDM and the PBP3 insert in Enterobacterales\",\"authors\":\"Yamuna Devi Bakthavatchalam , Fizaa Abdullah , Devishree Srinivasan , Sangeetha Nithiyanandam , Ayyanraj Neeravi , Poojah Shah , Nivedhana Subburaju , Subha Vajjiravelu Jaganathan , Rema Devi , Gita Nataraj , Binesh Lal Yesudason , Kamini Walia , Balaji Veeraraghavan\",\"doi\":\"10.1016/j.ijmmb.2024.100710\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><p>There is a high diversity of beta-lactamases in gram negative pathogens, making them difficult to treat. In the presence of OXA-1 and ampC, PTZ is no longer clinically relevant when treating Enterobacterales expressing ESBLs. Further, MBL infections are often treated with the combination of ceftazidime/avibactam with aztreonam. . It has recently been reported that NDM-expressing <em>E. coli</em> isolates co-harboring PBP3 insert develops resistance to this triple combination.</p></div><div><h3>Methods</h3><p>A pentaplex PCR is developed and validated to simultaneously detect bla<sub>CTX-M</sub>, bla<sub>OXA-1</sub>, bla<sub>CMY</sub>, bla<sub>NDM</sub>, and the PBP3 insert in whole genome sequenced <em>E. coli</em> and <em>K. pneumoniae</em> isolates. In addition, the isolates chosen for pentaplex PCR evaluation were tested for their minimum inhibitory concentrations (MICs) against piperacillin/tazobactam, cefoperazone/sulbactam (C/S), ertapenem, imipenem, meropenem, ceftazidime/avibactam, aztreonam/avibactam, cefepime/taniborbactam, and cefiderocol.</p></div><div><h3>Results</h3><p>The developed pentaplex PCR showed 100 % reproducibility with the antimicrobial resistance profile generated from whole genome sequenced data. PTZ and C/S are not effective against ESBL and/or OXA-1 expressing <em>E. coli</em> and <em>K. pneumoniae</em> isolates and do not offer any activity against CMY co-producers. Further, the combined effect of CMY, NDM and PBP3 inserts impacts aztreonam/avibactam activity and reduces the susceptibility to 40 % in <em>E. coli</em> isolates. While, aztreonam/avibactam showed potent activity against NDM-expressing <em>K. pneumoniae</em> isolates. Importantly, cefepime/taniborbactam and cefiderocol showed limited activity against NDM-expressing <em>E. coli</em> and <em>K. pneumoniae</em> isolates.</p></div><div><h3>Conclusion</h3><p>The pentaplex PCR was effective in detecting four beta-lactamases (bla<sub>CTX-M</sub>, bla<sub>OXA-1</sub>, bla<sub>CMY</sub>, bla<sub>NDM</sub>) as well as PBP3 inserts. It is expected that using pentaplex PCR as a diagnostic test for resistance detection in clinical practice will improve patient outcomes by providing prompt and targeted treatment.</p></div>\",\"PeriodicalId\":13284,\"journal\":{\"name\":\"Indian Journal of Medical Microbiology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.4000,\"publicationDate\":\"2024-08-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Indian Journal of Medical Microbiology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0255085724001853\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"IMMUNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Indian Journal of Medical Microbiology","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0255085724001853","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
Development and validation of a pentaplex PCR assay for rapid detection of blaCTX-M, blaOXA–1, blaCMY, blaNDM and the PBP3 insert in Enterobacterales
Background
There is a high diversity of beta-lactamases in gram negative pathogens, making them difficult to treat. In the presence of OXA-1 and ampC, PTZ is no longer clinically relevant when treating Enterobacterales expressing ESBLs. Further, MBL infections are often treated with the combination of ceftazidime/avibactam with aztreonam. . It has recently been reported that NDM-expressing E. coli isolates co-harboring PBP3 insert develops resistance to this triple combination.
Methods
A pentaplex PCR is developed and validated to simultaneously detect blaCTX-M, blaOXA-1, blaCMY, blaNDM, and the PBP3 insert in whole genome sequenced E. coli and K. pneumoniae isolates. In addition, the isolates chosen for pentaplex PCR evaluation were tested for their minimum inhibitory concentrations (MICs) against piperacillin/tazobactam, cefoperazone/sulbactam (C/S), ertapenem, imipenem, meropenem, ceftazidime/avibactam, aztreonam/avibactam, cefepime/taniborbactam, and cefiderocol.
Results
The developed pentaplex PCR showed 100 % reproducibility with the antimicrobial resistance profile generated from whole genome sequenced data. PTZ and C/S are not effective against ESBL and/or OXA-1 expressing E. coli and K. pneumoniae isolates and do not offer any activity against CMY co-producers. Further, the combined effect of CMY, NDM and PBP3 inserts impacts aztreonam/avibactam activity and reduces the susceptibility to 40 % in E. coli isolates. While, aztreonam/avibactam showed potent activity against NDM-expressing K. pneumoniae isolates. Importantly, cefepime/taniborbactam and cefiderocol showed limited activity against NDM-expressing E. coli and K. pneumoniae isolates.
Conclusion
The pentaplex PCR was effective in detecting four beta-lactamases (blaCTX-M, blaOXA-1, blaCMY, blaNDM) as well as PBP3 inserts. It is expected that using pentaplex PCR as a diagnostic test for resistance detection in clinical practice will improve patient outcomes by providing prompt and targeted treatment.
期刊介绍：
Manuscripts of high standard in the form of original research, multicentric studies, meta analysis, are accepted. Current reports can be submitted as brief communications. Case reports must include review of current literature, clinical details, outcome and follow up. Letters to the editor must be a comment on or pertain to a manuscript already published in the IJMM or in relation to preliminary communication of a larger study.
Review articles, Special Articles or Guest Editorials are accepted on invitation.
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