Lucas J Osborn, Lindsay Osborn, Irvin Ibarra-Flores, Marisol Garcia, Kaitlyn Perez, Ali Farhadiayoubloo, Melissa Mitrou, Cristina Costales, Jennifer Dien Bard
{"title":"早期生长分离株自动和人工微量肉汤稀释抗菌素敏感性试验的性能评价。","authors":"Lucas J Osborn, Lindsay Osborn, Irvin Ibarra-Flores, Marisol Garcia, Kaitlyn Perez, Ali Farhadiayoubloo, Melissa Mitrou, Cristina Costales, Jennifer Dien Bard","doi":"10.1128/jcm.00236-25","DOIUrl":null,"url":null,"abstract":"<p><p>Prolonged turnaround times (TAT) represent a major limitation to current automated susceptibility testing systems and manual susceptibility-testing methods such as broth microdilution. As a result, targeted therapy for patients may be delayed, portending suboptimal clinical outcomes. One contributing factor is the 18-24 h of incubation prior to antimicrobial-susceptibility testing (AST) recommended by the Clinical Laboratory Standards Institute (CLSI) and some automated AST manufacturers. This study evaluates the performance of AST by manual broth microdilution (Thermo Fisher Sensititre) and an automated AST system (BD Phoenix) on isolates incubated for 6 h (early growth AST, egAST) compared with 18-24 h (standard growth AST, sgAST). An initial proof-of-concept study conducted on gram-negative (<i>n</i> = 5) and gram-positive (<i>n</i> = 2) quality control strains incubated for 6 or 24 h prior to Sensititre and Phoenix demonstrated 100% essential agreement. Subsequently, we evaluated the performance of egAST on gram-positive (<i>n</i> = 49, Phoenix; <i>n</i> = 46 Sensititre) and gram-negative (<i>n</i> = 81 Phoenix; <i>n</i> = 61 Sensititre) patient-derived isolates with diverse resistance profiles compared with sgAST. In total, 1,666 organism-drug combinations were tested by Sensititre (560 gram-positive and 1,106 gram-negative) and 1,927 by Phoenix (409 gram-positive and 1,518 gram-negative). For <i>Enterobacterales</i>, egAST using Phoenix revealed 1.3% minor errors (MiE), 0.17% major errors (ME), and 1.1% very major errors (VME) compared with sgAST. Phoenix egAST performance for <i>Pseudomonas aeruginosa</i> revealed 0.18% MiE and no ME or VME. Similarly, egAST of <i>Enterobacterales</i> by Sensititre revealed 1.5% MiE and no ME or VME, whereas 2.8% MiE, 6.6% ME, and no VME were observed for <i>P. aeruginosa</i>. For <i>Staphylococcus</i> spp. and <i>Enterococcus</i> spp., there were no MiE, ME, or VME on the Phoenix system, whereas early growth Sensititre showed 3.14% MiE, 0.3% ME, and 2.99% VME for <i>Staphylococcus</i> spp. and 6.8% MiE, 0.09% ME, and no VME for <i>Enterococcus</i> spp. Taken together, these data suggest that egAST represents a viable strategy to reduce the prolonged incubation period currently recommended by CLSI and select automated AST manufacturer guidelines without incurring any additional costs while simultaneously maintaining high concordance with reference standard methods.IMPORTANCETraditional antimicrobial-susceptibility testing (AST) methods typically span several days from the time of organism isolation. The majority of this time is spent waiting for a cultured isolate to incubate up to 1 day prior to AST. There exists an unmet need to provide more rapid AST as various rapid methods have been shown to reduce exposure to broad-spectrum antibiotics that select for antimicrobial resistance, shorten hospital stays, and improve clinical outcomes. Simultaneously, there is a need to ensure that rapid AST approaches are readily implemented in the clinical microbiology laboratory, with little to no added financial burden. This study demonstrates a cost-considerate and practical approach to reduce AST turnaround times by up to 18 h through the use of early growth isolates in combination with two commercial AST systems. The findings from this study complement those of previous reports describing the largely acceptable performance of disk diffusion using early growth isolates.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0023625"},"PeriodicalIF":5.4000,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345138/pdf/","citationCount":"0","resultStr":"{\"title\":\"Performance evaluation of early growth isolates for automated and manual broth microdilution antimicrobial-susceptibility testing.\",\"authors\":\"Lucas J Osborn, Lindsay Osborn, Irvin Ibarra-Flores, Marisol Garcia, Kaitlyn Perez, Ali Farhadiayoubloo, Melissa Mitrou, Cristina Costales, Jennifer Dien Bard\",\"doi\":\"10.1128/jcm.00236-25\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Prolonged turnaround times (TAT) represent a major limitation to current automated susceptibility testing systems and manual susceptibility-testing methods such as broth microdilution. As a result, targeted therapy for patients may be delayed, portending suboptimal clinical outcomes. One contributing factor is the 18-24 h of incubation prior to antimicrobial-susceptibility testing (AST) recommended by the Clinical Laboratory Standards Institute (CLSI) and some automated AST manufacturers. This study evaluates the performance of AST by manual broth microdilution (Thermo Fisher Sensititre) and an automated AST system (BD Phoenix) on isolates incubated for 6 h (early growth AST, egAST) compared with 18-24 h (standard growth AST, sgAST). An initial proof-of-concept study conducted on gram-negative (<i>n</i> = 5) and gram-positive (<i>n</i> = 2) quality control strains incubated for 6 or 24 h prior to Sensititre and Phoenix demonstrated 100% essential agreement. Subsequently, we evaluated the performance of egAST on gram-positive (<i>n</i> = 49, Phoenix; <i>n</i> = 46 Sensititre) and gram-negative (<i>n</i> = 81 Phoenix; <i>n</i> = 61 Sensititre) patient-derived isolates with diverse resistance profiles compared with sgAST. In total, 1,666 organism-drug combinations were tested by Sensititre (560 gram-positive and 1,106 gram-negative) and 1,927 by Phoenix (409 gram-positive and 1,518 gram-negative). For <i>Enterobacterales</i>, egAST using Phoenix revealed 1.3% minor errors (MiE), 0.17% major errors (ME), and 1.1% very major errors (VME) compared with sgAST. Phoenix egAST performance for <i>Pseudomonas aeruginosa</i> revealed 0.18% MiE and no ME or VME. Similarly, egAST of <i>Enterobacterales</i> by Sensititre revealed 1.5% MiE and no ME or VME, whereas 2.8% MiE, 6.6% ME, and no VME were observed for <i>P. aeruginosa</i>. For <i>Staphylococcus</i> spp. and <i>Enterococcus</i> spp., there were no MiE, ME, or VME on the Phoenix system, whereas early growth Sensititre showed 3.14% MiE, 0.3% ME, and 2.99% VME for <i>Staphylococcus</i> spp. and 6.8% MiE, 0.09% ME, and no VME for <i>Enterococcus</i> spp. Taken together, these data suggest that egAST represents a viable strategy to reduce the prolonged incubation period currently recommended by CLSI and select automated AST manufacturer guidelines without incurring any additional costs while simultaneously maintaining high concordance with reference standard methods.IMPORTANCETraditional antimicrobial-susceptibility testing (AST) methods typically span several days from the time of organism isolation. The majority of this time is spent waiting for a cultured isolate to incubate up to 1 day prior to AST. There exists an unmet need to provide more rapid AST as various rapid methods have been shown to reduce exposure to broad-spectrum antibiotics that select for antimicrobial resistance, shorten hospital stays, and improve clinical outcomes. Simultaneously, there is a need to ensure that rapid AST approaches are readily implemented in the clinical microbiology laboratory, with little to no added financial burden. This study demonstrates a cost-considerate and practical approach to reduce AST turnaround times by up to 18 h through the use of early growth isolates in combination with two commercial AST systems. The findings from this study complement those of previous reports describing the largely acceptable performance of disk diffusion using early growth isolates.</p>\",\"PeriodicalId\":15511,\"journal\":{\"name\":\"Journal of Clinical Microbiology\",\"volume\":\" \",\"pages\":\"e0023625\"},\"PeriodicalIF\":5.4000,\"publicationDate\":\"2025-08-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345138/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Clinical Microbiology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1128/jcm.00236-25\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/6/30 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q1\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Clinical Microbiology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1128/jcm.00236-25","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/6/30 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Performance evaluation of early growth isolates for automated and manual broth microdilution antimicrobial-susceptibility testing.
Prolonged turnaround times (TAT) represent a major limitation to current automated susceptibility testing systems and manual susceptibility-testing methods such as broth microdilution. As a result, targeted therapy for patients may be delayed, portending suboptimal clinical outcomes. One contributing factor is the 18-24 h of incubation prior to antimicrobial-susceptibility testing (AST) recommended by the Clinical Laboratory Standards Institute (CLSI) and some automated AST manufacturers. This study evaluates the performance of AST by manual broth microdilution (Thermo Fisher Sensititre) and an automated AST system (BD Phoenix) on isolates incubated for 6 h (early growth AST, egAST) compared with 18-24 h (standard growth AST, sgAST). An initial proof-of-concept study conducted on gram-negative (n = 5) and gram-positive (n = 2) quality control strains incubated for 6 or 24 h prior to Sensititre and Phoenix demonstrated 100% essential agreement. Subsequently, we evaluated the performance of egAST on gram-positive (n = 49, Phoenix; n = 46 Sensititre) and gram-negative (n = 81 Phoenix; n = 61 Sensititre) patient-derived isolates with diverse resistance profiles compared with sgAST. In total, 1,666 organism-drug combinations were tested by Sensititre (560 gram-positive and 1,106 gram-negative) and 1,927 by Phoenix (409 gram-positive and 1,518 gram-negative). For Enterobacterales, egAST using Phoenix revealed 1.3% minor errors (MiE), 0.17% major errors (ME), and 1.1% very major errors (VME) compared with sgAST. Phoenix egAST performance for Pseudomonas aeruginosa revealed 0.18% MiE and no ME or VME. Similarly, egAST of Enterobacterales by Sensititre revealed 1.5% MiE and no ME or VME, whereas 2.8% MiE, 6.6% ME, and no VME were observed for P. aeruginosa. For Staphylococcus spp. and Enterococcus spp., there were no MiE, ME, or VME on the Phoenix system, whereas early growth Sensititre showed 3.14% MiE, 0.3% ME, and 2.99% VME for Staphylococcus spp. and 6.8% MiE, 0.09% ME, and no VME for Enterococcus spp. Taken together, these data suggest that egAST represents a viable strategy to reduce the prolonged incubation period currently recommended by CLSI and select automated AST manufacturer guidelines without incurring any additional costs while simultaneously maintaining high concordance with reference standard methods.IMPORTANCETraditional antimicrobial-susceptibility testing (AST) methods typically span several days from the time of organism isolation. The majority of this time is spent waiting for a cultured isolate to incubate up to 1 day prior to AST. There exists an unmet need to provide more rapid AST as various rapid methods have been shown to reduce exposure to broad-spectrum antibiotics that select for antimicrobial resistance, shorten hospital stays, and improve clinical outcomes. Simultaneously, there is a need to ensure that rapid AST approaches are readily implemented in the clinical microbiology laboratory, with little to no added financial burden. This study demonstrates a cost-considerate and practical approach to reduce AST turnaround times by up to 18 h through the use of early growth isolates in combination with two commercial AST systems. The findings from this study complement those of previous reports describing the largely acceptable performance of disk diffusion using early growth isolates.
期刊介绍:
The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.