Direct detection of extended-spectrum beta-lactamases (CTX-M) from blood cultures by LC-MS/MS bottom-up proteomics
Rapid bacterial species identification and antibiotic susceptibility testing in positive blood cultures have an important impact on the antibiotic treatment for patients. To identify extended-spectrum beta-lactamases (ESBL) directly in positive blood culture bottles, we developed a workflow of sapon...
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| Veröffentlicht in: | European journal of clinical microbiology & infectious diseases 2017-09, Vol.36 (9), p.1621-1628 |
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| container_title | European journal of clinical microbiology & infectious diseases |
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| creator | Fleurbaaij, F. Goessens, W. van Leeuwen, H. C. Kraakman, M. E. M. Bernards, S. T. Hensbergen, P. J. Kuijper, E. J. |
| description | Rapid bacterial species identification and antibiotic susceptibility testing in positive blood cultures have an important impact on the antibiotic treatment for patients. To identify extended-spectrum beta-lactamases (ESBL) directly in positive blood culture bottles, we developed a workflow of saponin extraction followed by a bottom-up proteomics approach using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The workflow was applied to positive blood cultures with
Escherichia coli
and
Klebsiella pneumoniae
collected prospectively in two academic hospitals over a 4-month period. Of 170 positive blood cultures, 22 (12.9%) contained ESBL-positive isolates based on standard susceptibility testing. Proteomic analysis identified CTX-M ESBLs in 95% of these isolates directly in positive blood cultures, whereas no false positives were found in the non-ESBL producing positive blood cultures. The results were confirmed by molecular characterisation of beta-lactamase genes. Based on this proof-of-concept study, we conclude that LC-MS/MS-based protein analysis can directly identify extended-spectrum beta lactamases in
E. coli
and
K. pneumoniae
positive blood cultures, and could be further developed for application in routine diagnostics. |
| doi_str_mv | 10.1007/s10096-017-2975-y |
| format | Article |
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Escherichia coli
and
Klebsiella pneumoniae
collected prospectively in two academic hospitals over a 4-month period. Of 170 positive blood cultures, 22 (12.9%) contained ESBL-positive isolates based on standard susceptibility testing. Proteomic analysis identified CTX-M ESBLs in 95% of these isolates directly in positive blood cultures, whereas no false positives were found in the non-ESBL producing positive blood cultures. The results were confirmed by molecular characterisation of beta-lactamase genes. Based on this proof-of-concept study, we conclude that LC-MS/MS-based protein analysis can directly identify extended-spectrum beta lactamases in
E. coli
and
K. pneumoniae
positive blood cultures, and could be further developed for application in routine diagnostics.</description><identifier>ISSN: 0934-9723</identifier><identifier>EISSN: 1435-4373</identifier><identifier>DOI: 10.1007/s10096-017-2975-y</identifier><identifier>PMID: 28397101</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Anti-Bacterial Agents - pharmacology ; Antibiotics ; Bacteria ; beta-Lactamases - chemistry ; beta-Lactamases - genetics ; beta-Lactamases - metabolism ; Biomarkers ; Biomedical and Life Sciences ; Biomedicine ; Blood ; Blood Culture ; Chromatography, Liquid ; E coli ; Escherichia coli - drug effects ; Escherichia coli - genetics ; Escherichia coli Infections - diagnosis ; Escherichia coli Infections - drug therapy ; Escherichia coli Infections - microbiology ; Faith healing ; Humans ; Internal Medicine ; Klebsiella ; Klebsiella Infections - diagnosis ; Klebsiella Infections - drug therapy ; Klebsiella Infections - microbiology ; Klebsiella pneumoniae ; Klebsiella pneumoniae - drug effects ; Klebsiella pneumoniae - genetics ; Liquid chromatography ; Mass spectrometry ; Mass spectroscopy ; Medical Microbiology ; Microbial Sensitivity Tests ; Original ; Original Article ; Prayer ; Prospective Studies ; Proteomics ; Proteomics - methods ; Sequence Analysis, DNA ; Workflow</subject><ispartof>European journal of clinical microbiology & infectious diseases, 2017-09, Vol.36 (9), p.1621-1628</ispartof><rights>The Author(s) 2017</rights><rights>European Journal of Clinical Microbiology & Infectious Diseases is a copyright of Springer, 2017.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c470t-1ccd793301eec08fdd345216cacfe4b5838aa25e91211a0abfe455eebfbd3adf3</citedby><cites>FETCH-LOGICAL-c470t-1ccd793301eec08fdd345216cacfe4b5838aa25e91211a0abfe455eebfbd3adf3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10096-017-2975-y$$EPDF$$P50$$Gspringer$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10096-017-2975-y$$EHTML$$P50$$Gspringer$$Hfree_for_read</linktohtml><link.rule.ids>230,314,776,780,881,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28397101$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fleurbaaij, F.</creatorcontrib><creatorcontrib>Goessens, W.</creatorcontrib><creatorcontrib>van Leeuwen, H. C.</creatorcontrib><creatorcontrib>Kraakman, M. E. M.</creatorcontrib><creatorcontrib>Bernards, S. T.</creatorcontrib><creatorcontrib>Hensbergen, P. J.</creatorcontrib><creatorcontrib>Kuijper, E. J.</creatorcontrib><title>Direct detection of extended-spectrum beta-lactamases (CTX-M) from blood cultures by LC-MS/MS bottom-up proteomics</title><title>European journal of clinical microbiology & infectious diseases</title><addtitle>Eur J Clin Microbiol Infect Dis</addtitle><addtitle>Eur J Clin Microbiol Infect Dis</addtitle><description>Rapid bacterial species identification and antibiotic susceptibility testing in positive blood cultures have an important impact on the antibiotic treatment for patients. To identify extended-spectrum beta-lactamases (ESBL) directly in positive blood culture bottles, we developed a workflow of saponin extraction followed by a bottom-up proteomics approach using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The workflow was applied to positive blood cultures with
Escherichia coli
and
Klebsiella pneumoniae
collected prospectively in two academic hospitals over a 4-month period. Of 170 positive blood cultures, 22 (12.9%) contained ESBL-positive isolates based on standard susceptibility testing. Proteomic analysis identified CTX-M ESBLs in 95% of these isolates directly in positive blood cultures, whereas no false positives were found in the non-ESBL producing positive blood cultures. The results were confirmed by molecular characterisation of beta-lactamase genes. Based on this proof-of-concept study, we conclude that LC-MS/MS-based protein analysis can directly identify extended-spectrum beta lactamases in
E. coli
and
K. pneumoniae
positive blood cultures, and could be further developed for application in routine diagnostics.</description><subject>Anti-Bacterial Agents - pharmacology</subject><subject>Antibiotics</subject><subject>Bacteria</subject><subject>beta-Lactamases - chemistry</subject><subject>beta-Lactamases - genetics</subject><subject>beta-Lactamases - metabolism</subject><subject>Biomarkers</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Blood</subject><subject>Blood Culture</subject><subject>Chromatography, Liquid</subject><subject>E coli</subject><subject>Escherichia coli - drug effects</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli Infections - diagnosis</subject><subject>Escherichia coli Infections - drug therapy</subject><subject>Escherichia coli Infections - microbiology</subject><subject>Faith healing</subject><subject>Humans</subject><subject>Internal Medicine</subject><subject>Klebsiella</subject><subject>Klebsiella Infections - diagnosis</subject><subject>Klebsiella Infections - drug therapy</subject><subject>Klebsiella Infections - microbiology</subject><subject>Klebsiella pneumoniae</subject><subject>Klebsiella pneumoniae - drug effects</subject><subject>Klebsiella pneumoniae - genetics</subject><subject>Liquid chromatography</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>Medical Microbiology</subject><subject>Microbial Sensitivity Tests</subject><subject>Original</subject><subject>Original Article</subject><subject>Prayer</subject><subject>Prospective Studies</subject><subject>Proteomics</subject><subject>Proteomics - methods</subject><subject>Sequence Analysis, DNA</subject><subject>Workflow</subject><issn>0934-9723</issn><issn>1435-4373</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp1kU1LHTEUhkOp1FvbH9BNCXRjF6n5mNxMNoJc-wX34kIL3YVMcsaOzEzGJCPef2_kWlHBTQ6c9zlvzuFF6BOj3xil6iiVVy8JZYpwrSTZvkELVglJKqHEW7SgWlREKy720fuUrmiZqZV6h_Z5LbRilC1QPO0iuIw95FK6MOLQYrjNMHrwJE2lGecBN5At6a3LdrAJEj5cXfwlm6-4jaGIfQgeu7nPcyxas8XrFdmcH23OcRNyDgOZJzzFkCEMnUsf0F5r-wQfH-oB-vPj-8XqF1mf_fy9OlkTVymaCXPOKy0EZQCO1q33opKcLZ11LVSNrEVtLZegGWfMUtuUrpQATdt4YX0rDtDxzneamwG8gzFH25spdoONWxNsZ54rY_fPXIYbI6WseK2KweGDQQzXM6Rshi456Hs7QpiTYXW9VJJTJgv65QV6FeY4lvMM01zpsriqCsV2lIshpQjt4zKMmvtEzS5RUxI194mabZn5_PSKx4n_ERaA74BUpPES4pOvX3W9A89erpM</recordid><startdate>20170901</startdate><enddate>20170901</enddate><creator>Fleurbaaij, F.</creator><creator>Goessens, W.</creator><creator>van Leeuwen, H. 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C.</au><au>Kraakman, M. E. M.</au><au>Bernards, S. T.</au><au>Hensbergen, P. J.</au><au>Kuijper, E. J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Direct detection of extended-spectrum beta-lactamases (CTX-M) from blood cultures by LC-MS/MS bottom-up proteomics</atitle><jtitle>European journal of clinical microbiology & infectious diseases</jtitle><stitle>Eur J Clin Microbiol Infect Dis</stitle><addtitle>Eur J Clin Microbiol Infect Dis</addtitle><date>2017-09-01</date><risdate>2017</risdate><volume>36</volume><issue>9</issue><spage>1621</spage><epage>1628</epage><pages>1621-1628</pages><issn>0934-9723</issn><eissn>1435-4373</eissn><abstract>Rapid bacterial species identification and antibiotic susceptibility testing in positive blood cultures have an important impact on the antibiotic treatment for patients. To identify extended-spectrum beta-lactamases (ESBL) directly in positive blood culture bottles, we developed a workflow of saponin extraction followed by a bottom-up proteomics approach using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The workflow was applied to positive blood cultures with
Escherichia coli
and
Klebsiella pneumoniae
collected prospectively in two academic hospitals over a 4-month period. Of 170 positive blood cultures, 22 (12.9%) contained ESBL-positive isolates based on standard susceptibility testing. Proteomic analysis identified CTX-M ESBLs in 95% of these isolates directly in positive blood cultures, whereas no false positives were found in the non-ESBL producing positive blood cultures. The results were confirmed by molecular characterisation of beta-lactamase genes. Based on this proof-of-concept study, we conclude that LC-MS/MS-based protein analysis can directly identify extended-spectrum beta lactamases in
E. coli
and
K. pneumoniae
positive blood cultures, and could be further developed for application in routine diagnostics.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>28397101</pmid><doi>10.1007/s10096-017-2975-y</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | European journal of clinical microbiology & infectious diseases, 2017-09, Vol.36 (9), p.1621-1628 |
| issn | 0934-9723 1435-4373 |
| language | eng |
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| subjects | Anti-Bacterial Agents - pharmacology Antibiotics Bacteria beta-Lactamases - chemistry beta-Lactamases - genetics beta-Lactamases - metabolism Biomarkers Biomedical and Life Sciences Biomedicine Blood Blood Culture Chromatography, Liquid E coli Escherichia coli - drug effects Escherichia coli - genetics Escherichia coli Infections - diagnosis Escherichia coli Infections - drug therapy Escherichia coli Infections - microbiology Faith healing Humans Internal Medicine Klebsiella Klebsiella Infections - diagnosis Klebsiella Infections - drug therapy Klebsiella Infections - microbiology Klebsiella pneumoniae Klebsiella pneumoniae - drug effects Klebsiella pneumoniae - genetics Liquid chromatography Mass spectrometry Mass spectroscopy Medical Microbiology Microbial Sensitivity Tests Original Original Article Prayer Prospective Studies Proteomics Proteomics - methods Sequence Analysis, DNA Workflow |
| title | Direct detection of extended-spectrum beta-lactamases (CTX-M) from blood cultures by LC-MS/MS bottom-up proteomics |
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