{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T16:33:46Z","timestamp":1792600426381,"version":"4.5.0"},"reference-count":27,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,4,28]],"date-time":"2021-04-28T00:00:00Z","timestamp":1619568000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,4,28]],"date-time":"2021-04-28T00:00:00Z","timestamp":1619568000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["CP-151952"],"award-info":[{"award-number":["CP-151952"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["CP-151952"],"award-info":[{"award-number":["CP-151952"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Antimicrobial susceptibility testing (AST) is essential for detecting resistance in <jats:italic>Pseudomonas aeruginosa<\/jats:italic> and other bacterial pathogens. Here we evaluated the performance of broth microdilution (BMD) panels created using a semi-automated liquid handler, the D300e Digital Dispenser (Tecan Group Ltd., CH) that relies on inkjet printing technology. Microtitre panels (96-well) containing nine twofold dilutions of 12 antimicrobials from five classes (\u03b2-lactams, \u03b2-lactam\/\u03b2-lactamase inhibitors, aminoglycosides, fluoroquinolones, polymyxins) were prepared in parallel using the D300e Digital Dispenser and standard methods described by CLSI\/ISO. To assess performance, panels were challenged with three well characterized quality control organisms and 100 clinical <jats:italic>P. aeruginosa<\/jats:italic> isolates. Traditional agreement and error measures were used for evaluation. Essential (EA) and categorical (CA) agreements were 92.7% and 98.0% respectively for <jats:italic>P. aeruginosa<\/jats:italic> isolates with evaluable on-scale results. The majority of minor errors that fell outside acceptable EA parameters (\u2265\u2009\u00b1\u20091 dilution, 1.9%) were seen with aztreonam (5%) and ceftazidime (4%), however all antimicrobials displayed acceptable performance in this situation. Differences in MIC were often log<jats:sub>2<\/jats:sub> dilution lower for D300e dispensed panels. Major and very major errors were noted for aztreonam (2.6%) and cefepime (1.7%) respectively. The variable performance of D300e panels suggests that further testing is required to confirm their diagnostic utility for <jats:italic>P. aeruginosa<\/jats:italic>.<\/jats:p>","DOI":"10.1038\/s41598-021-88423-0","type":"journal-article","created":{"date-parts":[[2021,4,28]],"date-time":"2021-04-28T10:04:55Z","timestamp":1619604295000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Evaluation of digital dispense-assisted broth microdilution antimicrobial susceptibility testing for Pseudomonas aeruginosa isolates"],"prefix":"10.1038","volume":"11","author":[{"given":"Shawn T.","family":"Clark","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick J.","family":"Stapleton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pauline W.","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yvonne C. W.","family":"Yau","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valerie J.","family":"Waters","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David M.","family":"Hwang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David S.","family":"Guttman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,4,28]]},"reference":[{"key":"88423_CR1","doi-asserted-by":"publisher","first-page":"619","DOI":"10.1093\/jac\/dks439","volume":"68","author":"C Juan","year":"2013","unstructured":"Juan, C. et al. Challenges for accurate susceptibility testing, detection and interpretation of \u03b2-lactam resistance phenotypes in Pseudomonas aeruginosa: Results from a Spanish multicentre study. J. Antimicrob. Chemother. 68, 619\u2013630 (2013).","journal-title":"J. Antimicrob. Chemother."},{"key":"88423_CR2","unstructured":"ISO. Susceptibility testing of infectious agents and evaluation of performance of antimicrobial susceptibility test devices\u2014Part 1: Broth micro-dilution reference method for testing the in vitro activity of antimicrobial agents against rapidly growing aerobic bacteria involved in infectious diseases. International Standard Document 20776-1, 2nd ed (2019)."},{"key":"88423_CR3","unstructured":"Clinical and Laboratory Standards Institute. Performance Standards for antimicrobial susceptibility testing. 30th ed. CLSI Supplement M100. (Clinical and Laboratory Standards Institute, 2020)."},{"key":"88423_CR4","doi-asserted-by":"publisher","first-page":"1749","DOI":"10.1086\/647952","volume":"49","author":"JH Jorgensen","year":"2009","unstructured":"Jorgensen, J. H. & Ferraro, M. J. Antimicrobial susceptibility testing: A review of general principles and contemporary practices. Clin. Infect. Dis. 49, 1749\u20131755 (2009).","journal-title":"Clin. Infect. Dis."},{"key":"88423_CR5","doi-asserted-by":"publisher","first-page":"2288","DOI":"10.1128\/JCM.00932-16","volume":"54","author":"KP Smith","year":"2016","unstructured":"Smith, K. P. & Kirby, J. E. Verification of an automated, digital dispensing platform for at-Will Broth microdilution-based antimicrobial susceptibility testing. J. Clin. Microbiol. 54, 2288\u20132293 (2016).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR6","doi-asserted-by":"publisher","first-page":"470","DOI":"10.1128\/JCM.02128-16","volume":"55","author":"KP Smith","year":"2017","unstructured":"Smith, K. P., Brennan-Krohn, T., Weir, S. & Kirby, J. E. Improved accuracy of cefepime susceptibility testing for extended-spectrum-beta-lactamase-producing enterobacteriaceae with an on-demand digital dispensing method. J. Clin. Microbiol. 55, 470\u2013478 (2017).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR7","doi-asserted-by":"publisher","first-page":"2304","DOI":"10.1128\/JCM.00511-17","volume":"55","author":"T Brennan-Krohn","year":"2017","unstructured":"Brennan-Krohn, T., Smith, K. P. & Kirby, J. E. The poisoned well: Enhancing the predictive value of antimicrobial susceptibility testing in the era of multidrug resistance. J. Clin. Microbiol. 55, 2304\u20132308 (2017).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR8","doi-asserted-by":"publisher","first-page":"2775","DOI":"10.1093\/jac\/dkx241","volume":"72","author":"T Brennan-Krohn","year":"2017","unstructured":"Brennan-Krohn, T., Truelson, K. A., Smith, K. P. & Kirby, J. E. Screening for synergistic activity of antimicrobial combinations against carbapenem-resistant Enterobacteriaceae using inkjet printer-based technology. J. Antimicrob. Chemother. 72, 2775\u20132781 (2017).","journal-title":"J. Antimicrob. Chemother."},{"key":"88423_CR9","first-page":"e01944","volume":"58","author":"E Ransom","year":"2020","unstructured":"Ransom, E. et al. Validation of aztreonam-avibactam susceptibility testing using digitally dispensed custom panels. J. Clin. Microbiol. 58, e01944-e2019 (2020).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR10","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1177\/2211068213491094","volume":"18","author":"RE Jones","year":"2013","unstructured":"Jones, R. E., Zheng, W., McKew, J. C. & Chen, C. Z. An alternative direct compound dispensing method using the HP D300 digital dispenser. J. Lab. Autom. 18, 367\u2013374 (2013).","journal-title":"J. Lab. Autom."},{"key":"88423_CR11","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1016\/j.jim.2016.12.003","volume":"442","author":"C Roselle","year":"2017","unstructured":"Roselle, C. et al. Evaluation of a digital dispenser for direct curve dilutions in a vaccine potency assay. J. Immunol. Methods 442, 20\u201328 (2017).","journal-title":"J. Immunol. Methods"},{"key":"88423_CR12","doi-asserted-by":"publisher","first-page":"1215","DOI":"10.1002\/rcm.6894","volume":"28","author":"D Liao","year":"2014","unstructured":"Liao, D., Chen, S., Paton, M. & Qian, M. G. Automated non-stepwise preparation of bioanalytical calibration standards and quality controls using an ultra-low volume digitizing liquid dispenser. Rapid Commun. Mass Spectrom. 28, 1215\u20131220 (2014).","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"88423_CR13","unstructured":"Clinical and Laboratory Standards Institute. Verification of Commercial Microbial Identification and Antimicrobial Susceptibility Testing Systems, 1st ed. CLSI M52 (Clinical and Laboratory Standards Institute, Wayne, Pennsylvania, 2015)."},{"key":"88423_CR14","unstructured":"Clinical and Laboratory Standards Institute. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, 11th ed. CLSI standard M07 (Clinical and Laboratory Standards Institute, Wayne, Pennsylvania, 2018)."},{"key":"88423_CR15","volume-title":"Cumitech 31A: Verification and Validation of Procedures in the Clinical Microbiology Laboratory Coordinating SE Sharp","author":"RB Clark","year":"2009","unstructured":"Clark, R. B., Lewinski, M. A., Loeffelholz, M. J. & Tibbetts, R. J. Cumitech 31A: Verification and Validation of Procedures in the Clinical Microbiology Laboratory (eds Sharp, S. E. & Clark, R. B.) (ASM Press, Washington, DC, 2009)."},{"key":"88423_CR16","first-page":"e01934","volume":"56","author":"RM Humphries","year":"2018","unstructured":"Humphries, R. M. et al. CLSI Methods development and standardization working group best practices for evaluation of antimicrobial susceptibility tests. J. Clin. Microbiol. 56, e01934-e2017 (2018).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR17","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1111\/j.1600-0463.2009.02436.x","volume":"117","author":"WS Hsieh","year":"2009","unstructured":"Hsieh, W. S., Sung, L. L., Tsai, K. C. & Ho, H. T. Evaluation of the VITEK 2 cards for identification and antimicrobial susceptibility testing of non-glucose fermenting Gram-negative bacilli. APMIS 117, 241\u2013247 (2009).","journal-title":"APMIS"},{"key":"88423_CR18","doi-asserted-by":"publisher","first-page":"450","DOI":"10.1128\/JCM.01859-16","volume":"55","author":"AM Bobenchik","year":"2017","unstructured":"Bobenchik, A. M., Deak, E., Hindler, J. A., Charlton, C. L. & Humphries, R. M. Performance of Vitek 2 for antimicrobial susceptibility testing of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomas maltophilia with Vitek 2 (2009 FDA) and CLSI M100S 26th edition breakpoints. J. Clin. Microbiol. 55, 450\u2013456 (2017).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR19","doi-asserted-by":"crossref","first-page":"e01042","DOI":"10.1128\/JCM.01042-19","volume":"58","author":"S Garc\u00eda-Fernandez","year":"2020","unstructured":"Garc\u00eda-Fernandez, S. et al. Multicenter evaluation of the new Etest gradient diffusion method for piperacillin-tazobactam susceptibility testing of Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter baumannii complex. J. Clin. Microbiol. 58, e01042-e1119 (2020).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR20","doi-asserted-by":"publisher","first-page":"625","DOI":"10.1099\/jmm.0.005587-0","volume":"58","author":"E Torres","year":"2009","unstructured":"Torres, E., Villanueva, R. & Bou, G. Comparison of different methods of determining B-lactam susceptibility in clinical strains of Pseudomonas aeruginosa. J. Med. Microbiol. 58, 625\u2013629 (2009).","journal-title":"J. Med. Microbiol."},{"key":"88423_CR21","doi-asserted-by":"publisher","first-page":"1339","DOI":"10.1128\/JCM.01716-06","volume":"45","author":"S Juretschko","year":"2007","unstructured":"Juretschko, S., LaBombardi, V. J., Lerner, S. A., Schreckenberger, P. C. & Pseudomonas AST Study Group. Accuracies of \u03b2-lactam susceptibility test results for Pseudomonas aeruginosa with four automated systems (BD Phoenix, MicroScan WalkAway, Vitek, and Vitek 2). J. Clin. Microbiol. 45, 1339\u20131342 (2007).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR22","doi-asserted-by":"publisher","first-page":"1678","DOI":"10.1128\/JCM.03385-12","volume":"51","author":"JA Hindler","year":"2013","unstructured":"Hindler, J. A. & Humphries, R. M. Colistin MIC variability by method for contemporary clinical isolates of multi-drug resistant Gram-negative bacilli. J. Clin. Microbiol. 51, 1678\u20131684 (2013).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR23","doi-asserted-by":"publisher","first-page":"1416","DOI":"10.1093\/jac\/dkq131","volume":"65","author":"SM Moskowitz","year":"2010","unstructured":"Moskowitz, S. M. et al. Colistin susceptibility testing: Evaluation of reliability for cystic fibrosis isolates of Pseudomonas aeruginosa and Stenotrophomonas maltophilia. J. Antimicrob. Chemother. 65, 1416\u20131423 (2010).","journal-title":"J. Antimicrob. Chemother."},{"key":"88423_CR24","doi-asserted-by":"publisher","first-page":"645","DOI":"10.1093\/jac\/dky477","volume":"74","author":"AJ Denisuik","year":"2019","unstructured":"Denisuik, A. J. et al. Antimicrobial-resistant pathogens in Canadian ICUs: Results of the CANWARD 2007 to 2016 study. J. Antimicrob. Chemother. 74, 645\u2013653 (2019).","journal-title":"J. Antimicrob. Chemother."},{"key":"88423_CR25","first-page":"e00203","volume":"57","author":"RM Humphries","year":"2019","unstructured":"Humphries, R. M., Abbott, A. N. & Hindler, J. A. Understanding and addressing CLSI breakpoint revisions: A primer for clinical laboratories. J. Clin. Microbiol. 57, e00203-e219 (2019).","journal-title":"J. Clin. Microbiol."},{"key":"88423_CR26","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1038\/s41579-018-0098-9","volume":"17","author":"A van Belkum","year":"2019","unstructured":"van Belkum, A. et al. Developmental roadmap for antimicrobial susceptibility testing systems. Nat. Rev. Microbiol. 17, 51\u201362 (2019).","journal-title":"Nat. Rev. Microbiol."},{"key":"88423_CR27","doi-asserted-by":"publisher","first-page":"e00873","DOI":"10.1128\/AAC.00873-18","volume":"62","author":"T Brennan-Krohn","year":"2018","unstructured":"Brennan-Krohn, T., Pironti, A. & Kirby, J. E. Synergistic activity of colistin-containing combinations against colistin-resistant enterobacteriaceae. Antimicrob. Agents Chemother. 62, e00873-e918 (2018).","journal-title":"Antimicrob. Agents Chemother."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-88423-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-88423-0","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-88423-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,3]],"date-time":"2022-12-03T07:23:43Z","timestamp":1670052223000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-88423-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,28]]},"references-count":27,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["88423"],"URL":"https:\/\/doi.org\/10.1038\/s41598-021-88423-0","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,28]]},"assertion":[{"value":"21 January 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 April 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"28 April 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"9157"}}