Antibiotic susceptibility testing of grown blood cultures by combining culture and real-time polymerase chain reaction is rapid and effective
Early administration of appropriate antibiotic therapy in bacteraemia patients dramatically reduces mortality. A new method for RApid Molecular Antibiotic Susceptibility Testing (RAMAST) that can be applied directly to positive blood cultures was developed and evaluated. Growth curves and antibiotic...
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| Veröffentlicht in: | PloS one 2011-12, Vol.6 (12), p.e27689-e27689 |
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| creator | Beuving, Judith Verbon, Annelies Gronthoud, Firza A Stobberingh, Ellen E Wolffs, Petra F G |
| description | Early administration of appropriate antibiotic therapy in bacteraemia patients dramatically reduces mortality. A new method for RApid Molecular Antibiotic Susceptibility Testing (RAMAST) that can be applied directly to positive blood cultures was developed and evaluated.
Growth curves and antibiotic susceptibility of blood culture isolates (Staphylococcus aureus, enterococci and (facultative) aerobic gram-negative rods) were determined by incubating diluted blood cultures with and without antibiotics, followed by a quantitative universal 16S PCR to detect the presence or absence of growth. Testing 114 positive blood cultures, RAMAST showed an agreement with microbroth dilution of 96.7% for gram-negative rods, with a minor error (false-susceptibility with a intermediate resistant strain) rate of 1.9%, a major error (false resistance) rate of 0.8% and a very major error (false susceptibility) rate of 0.6%. Agreement for S. aureus was 97.9%, with a very major error rate of 2.1%. Enterococcus species showed 95.0% agreement, with a major error rate of 5.0%. These agreements are comparable with those of the Phoenix system. Starting from a positive blood culture, the test was completed within 9 hours.
This new rapid method for antibiotic susceptibility testing can potentially provide accurate results for most relevant bacteria commonly isolated from positive blood cultures in less time than routine methods. |
| doi_str_mv | 10.1371/journal.pone.0027689 |
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Growth curves and antibiotic susceptibility of blood culture isolates (Staphylococcus aureus, enterococci and (facultative) aerobic gram-negative rods) were determined by incubating diluted blood cultures with and without antibiotics, followed by a quantitative universal 16S PCR to detect the presence or absence of growth. Testing 114 positive blood cultures, RAMAST showed an agreement with microbroth dilution of 96.7% for gram-negative rods, with a minor error (false-susceptibility with a intermediate resistant strain) rate of 1.9%, a major error (false resistance) rate of 0.8% and a very major error (false susceptibility) rate of 0.6%. Agreement for S. aureus was 97.9%, with a very major error rate of 2.1%. Enterococcus species showed 95.0% agreement, with a major error rate of 5.0%. These agreements are comparable with those of the Phoenix system. Starting from a positive blood culture, the test was completed within 9 hours.
This new rapid method for antibiotic susceptibility testing can potentially provide accurate results for most relevant bacteria commonly isolated from positive blood cultures in less time than routine methods.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0027689</identifier><identifier>PMID: 22194790</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Anti-Bacterial Agents - pharmacology ; Antibiotics ; Bacteremia ; Bacteremia - blood ; Bacteria ; Bacteria - drug effects ; Bacteria - growth & development ; Biology ; Blood ; Blood culture ; Blood tests ; Cell Culture Techniques - methods ; Clinical outcomes ; Culture ; Dilution ; Enterobacteriaceae ; Errors ; Ethics ; Growth curves ; Health aspects ; Humans ; Identification ; Kinetics ; Laboratories ; Mass spectrometry ; Medical research ; Medicine ; Microbial Sensitivity Tests - methods ; Microbiology ; Mortality ; Physics ; Polymerase chain reaction ; Pseudomonas aeruginosa ; Pseudomonas aeruginosa - drug effects ; Pseudomonas aeruginosa - growth & development ; Pseudomonas aeruginosa - isolation & purification ; Public health ; Real-Time Polymerase Chain Reaction - methods ; Rods ; Scientific imaging ; Sepsis ; Staphylococcus aureus ; Staphylococcus aureus - drug effects ; Staphylococcus aureus - growth & development ; Staphylococcus aureus - isolation & purification ; Test procedures</subject><ispartof>PloS one, 2011-12, Vol.6 (12), p.e27689-e27689</ispartof><rights>COPYRIGHT 2011 Public Library of Science</rights><rights>2011 Beuving et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>Beuving et al. 2011</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c691t-1eabf4d70b008f2b8fa20b95275623009fe8938a504a3e3dfd393824c697c8f23</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3237415/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3237415/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2095,2914,23846,27903,27904,53770,53772,79347,79348</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22194790$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Kaufmann, Gunnar F.</contributor><creatorcontrib>Beuving, Judith</creatorcontrib><creatorcontrib>Verbon, Annelies</creatorcontrib><creatorcontrib>Gronthoud, Firza A</creatorcontrib><creatorcontrib>Stobberingh, Ellen E</creatorcontrib><creatorcontrib>Wolffs, Petra F G</creatorcontrib><title>Antibiotic susceptibility testing of grown blood cultures by combining culture and real-time polymerase chain reaction is rapid and effective</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Early administration of appropriate antibiotic therapy in bacteraemia patients dramatically reduces mortality. A new method for RApid Molecular Antibiotic Susceptibility Testing (RAMAST) that can be applied directly to positive blood cultures was developed and evaluated.
Growth curves and antibiotic susceptibility of blood culture isolates (Staphylococcus aureus, enterococci and (facultative) aerobic gram-negative rods) were determined by incubating diluted blood cultures with and without antibiotics, followed by a quantitative universal 16S PCR to detect the presence or absence of growth. Testing 114 positive blood cultures, RAMAST showed an agreement with microbroth dilution of 96.7% for gram-negative rods, with a minor error (false-susceptibility with a intermediate resistant strain) rate of 1.9%, a major error (false resistance) rate of 0.8% and a very major error (false susceptibility) rate of 0.6%. Agreement for S. aureus was 97.9%, with a very major error rate of 2.1%. Enterococcus species showed 95.0% agreement, with a major error rate of 5.0%. These agreements are comparable with those of the Phoenix system. Starting from a positive blood culture, the test was completed within 9 hours.
This new rapid method for antibiotic susceptibility testing can potentially provide accurate results for most relevant bacteria commonly isolated from positive blood cultures in less time than routine methods.</description><subject>Anti-Bacterial Agents - pharmacology</subject><subject>Antibiotics</subject><subject>Bacteremia</subject><subject>Bacteremia - blood</subject><subject>Bacteria</subject><subject>Bacteria - drug effects</subject><subject>Bacteria - growth & development</subject><subject>Biology</subject><subject>Blood</subject><subject>Blood culture</subject><subject>Blood tests</subject><subject>Cell Culture Techniques - methods</subject><subject>Clinical outcomes</subject><subject>Culture</subject><subject>Dilution</subject><subject>Enterobacteriaceae</subject><subject>Errors</subject><subject>Ethics</subject><subject>Growth curves</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Identification</subject><subject>Kinetics</subject><subject>Laboratories</subject><subject>Mass spectrometry</subject><subject>Medical research</subject><subject>Medicine</subject><subject>Microbial Sensitivity Tests - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Beuving, Judith</au><au>Verbon, Annelies</au><au>Gronthoud, Firza A</au><au>Stobberingh, Ellen E</au><au>Wolffs, Petra F G</au><au>Kaufmann, Gunnar F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Antibiotic susceptibility testing of grown blood cultures by combining culture and real-time polymerase chain reaction is rapid and effective</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2011-12-14</date><risdate>2011</risdate><volume>6</volume><issue>12</issue><spage>e27689</spage><epage>e27689</epage><pages>e27689-e27689</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Early administration of appropriate antibiotic therapy in bacteraemia patients dramatically reduces mortality. A new method for RApid Molecular Antibiotic Susceptibility Testing (RAMAST) that can be applied directly to positive blood cultures was developed and evaluated.
Growth curves and antibiotic susceptibility of blood culture isolates (Staphylococcus aureus, enterococci and (facultative) aerobic gram-negative rods) were determined by incubating diluted blood cultures with and without antibiotics, followed by a quantitative universal 16S PCR to detect the presence or absence of growth. Testing 114 positive blood cultures, RAMAST showed an agreement with microbroth dilution of 96.7% for gram-negative rods, with a minor error (false-susceptibility with a intermediate resistant strain) rate of 1.9%, a major error (false resistance) rate of 0.8% and a very major error (false susceptibility) rate of 0.6%. Agreement for S. aureus was 97.9%, with a very major error rate of 2.1%. Enterococcus species showed 95.0% agreement, with a major error rate of 5.0%. These agreements are comparable with those of the Phoenix system. Starting from a positive blood culture, the test was completed within 9 hours.
This new rapid method for antibiotic susceptibility testing can potentially provide accurate results for most relevant bacteria commonly isolated from positive blood cultures in less time than routine methods.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>22194790</pmid><doi>10.1371/journal.pone.0027689</doi><tpages>e27689</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS); EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Anti-Bacterial Agents - pharmacology Antibiotics Bacteremia Bacteremia - blood Bacteria Bacteria - drug effects Bacteria - growth & development Biology Blood Blood culture Blood tests Cell Culture Techniques - methods Clinical outcomes Culture Dilution Enterobacteriaceae Errors Ethics Growth curves Health aspects Humans Identification Kinetics Laboratories Mass spectrometry Medical research Medicine Microbial Sensitivity Tests - methods Microbiology Mortality Physics Polymerase chain reaction Pseudomonas aeruginosa Pseudomonas aeruginosa - drug effects Pseudomonas aeruginosa - growth & development Pseudomonas aeruginosa - isolation & purification Public health Real-Time Polymerase Chain Reaction - methods Rods Scientific imaging Sepsis Staphylococcus aureus Staphylococcus aureus - drug effects Staphylococcus aureus - growth & development Staphylococcus aureus - isolation & purification Test procedures |
| title | Antibiotic susceptibility testing of grown blood cultures by combining culture and real-time polymerase chain reaction is rapid and effective |
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