Application of total laboratory automation for direct rapid antimicrobial susceptibility testing from Enterobacterales-containing blood cultures_West China Medical Journal

Authors：

ZHANG Weili ^1,2,3 , SHU Ling ^1,2,3 , LIAO Quanfeng ^1,2,3 , WU Siying ^1,2,3 , LIU Ya ^1,2,3 , DENG Jin ^1,2,3 , MA Ying ^1,2,3 , KANG Mei ^1,2,3 , XIE Yi ^1,2,3 ,  XIAO Yuling ^1,2,3

1. Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China;
2. Clinical Laboratory Medicine Research Center, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China;
3. Sichuan Clinical Research Center for Laboratory Medicine, Chengdu, Sichuan 610041, P. R. China;

Corresponding?author：

XIAO Yuling, Email: xieyi@scu.edu.cn

Keywords：

Blood cultures; Enterobacterales; rapid antimicrobial susceptibility testing; total laboratory automation; Clinical and Laboratory Standards Institute; European Committee on Antimicrobial Susceptibility Testing

DOI：

10.7507/1002-0179.202508022

Video：

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Objective To perform rapid antimicrobial susceptibility testing (RAST) on positive blood cultures of Enterobacterales using a total laboratory automation (TLA) system following both Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) standards, and to evaluate the two RAST methods. Methods Positive blood culture bottles growing Enterobacterales [54 Escherichia coli (E. coli) and 60 Klebsiella pneumoniae (K. pneumoniae) isolates] were collected at West China Hospital of Sichuan University between April and August 2022. CLSI RAST (8 and 16 h) and EUCAST RAST (4, 6, and 8 h) were performed using the TLA system, and results were compared with Vitek 2 antimicrobial susceptibility testing. Results CLSI RAST demonstrated lower categorical agreement with Vitek 2 (E. coli: 66.7% at 8 h, 81.9% at 16 h; K. pneumoniae: 72.8% at 8 h, 84.0% at 16 h) and tended to overcall resistance. EUCAST RAST showed increasing zone readability over time and high categorical agreement with Vitek 2 (E. coli: 97.1%, 96.2%, and 96.1% at 4, 6, and 8 h, respectively; K. pneumoniae: 96.1%, 97.1%, and 97.9% at 4, 6, and 8 h, respectively), as well as low error rates (major errors <3%). With the exception of ciprofloxacin, both the readability and categorical agreement for each antimicrobial agent against E. coli at 8 h were favorable (>90%). Similarly, for K. pneumoniae, the readability and categorical agreement for each antimicrobial agent at 8 h also demonstrated good performance (>90%). Conclusions According to CLSI-M52 criteria, EUCAST RAST at 4, 6, and 8 h shows equivalent performance to Vitek 2. Except for ciprofloxacin against E. coli, all antimicrobials tested at 8 h exhibit good readability and categorical agreement (>90%). CLSI RAST has lower categorical agreement and a higher tendency to interpret isolates as resistant.

Citation： ZHANG Weili, SHU Ling, LIAO Quanfeng, WU Siying, LIU Ya, DENG Jin, MA Ying, KANG Mei, XIE Yi, XIAO Yuling. Application of total laboratory automation for direct rapid antimicrobial susceptibility testing from Enterobacterales-containing blood cultures. West China Medical Journal, 2026, 41(5): 765-772. doi: 10.7507/1002-0179.202508022 Copy

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2.	Rudd KE, Kissoon N, Limmathurotsakul D, et al. The global burden of sepsis: barriers and potential solutions. Crit Care, 2018, 22(1): 232.
3.	Timsit JF, Ruppé E, Barbier F, et al. Bloodstream infections in critically ill patients: an expert statement. Intensive Care Med, 2020, 46(2): 266-284.
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8.	Herroelen PH, Heestermans R, Emmerechts K, et al. Validation of rapid antimicrobial susceptibility testing directly from blood cultures using WASPLab^?, including Colibrí^TM and Radian^? in-line carousel. Eur J Clin Microbiol Infect Dis, 2022, 41(5): 733-739.
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12.	Bianco G, Boattini M, Comini S, et al. Rapid determination of ceftazidime/avibactam susceptibility of carbapenemase-producing Enterobacterales directly from blood cultures: a comparative evaluation of EUCAST disc diffusion RAST and direct Etest^? RAST. J Antimicrob Chemother, 2022, 77(6): 1670-1675.
13.	Berinson B, Olearo F, Both A, et al. EUCAST rapid antimicrobial susceptibility testing (RAST): analytical performance and impact on patient management. J Antimicrob Chemother, 2021, 76(5): 1332-1338.
14.	Bin Najeeb MA, Gupta A, Purwar S, et al. Implementing EUCAST rapid antimicrobial susceptibility testing method for sepsis: lessons learned in a tertiary care center. J Infect Dev Ctries, 2021, 15(6): 833-839.
15.	Cherkaoui A, Schorderet D, Azam N, et al. Fully automated EUCAST rapid antimicrobial susceptibility testing (RAST) from positive blood cultures: diagnostic accuracy and implementation. J Clin Microbiol, 2022, 60(10): e0089822.
16.	Tay?i MR, ?entürk G?, ?ali?kan E, et al. Implementation of the EUCAST rapid antimicrobial susceptibility test (RAST) directly from positive blood culture bottles without the advanced identification systems. J Antimicrob Chemother, 2022, 77(4): 1020-1026.
17.	Jonasson E, Matuschek E, Kahlmeter G. The EUCAST rapid disc diffusion method for antimicrobial susceptibility testing directly from positive blood culture bottles. J Antimicrob Chemother, 2020, 75(4): 968-978.
18.	Bianco G, Lombardo D, Ricciardelli G, et al. Multicentre evaluation of the EUCAST rapid antimicrobial susceptibility testing (RAST) extending analysis to 16-20 hours reading time. Antibiotics (Basel), 2022, 11(10): 1404.
19.	Ekwall-Larson A, Fr?ding I, Mert B, et al. Analytical performance and potential clinical utility of EUCAST rapid antimicrobial susceptibility testing in blood cultures after four hours of incubation. Microbiol Spectr, 2023, 11(2): e0500122.
20.	Shan Y, Hu B, Guo W, et al. Evaluation of the EUCAST rapid antimicrobial susceptibility test for Enterobacterales-containing blood cultures in China. J Clin Microbiol, 2022, 60(4): e0255921.
21.	Martins A, Wink P, Pereira D, et al. Rapid antimicrobial susceptibility of Enterobacteriaceae by disk diffusion directly from blood culture bottles using the EUCAST RAST breakpoints. J Glob Antimicrob Resist, 2020, 22: 637-642.
22.	Valentin T, Koenig E, Prattes J, et al. Implementation of rapid antimicrobial susceptibility testing combined with routine infectious disease bedside consultation in clinical practice (RAST-ID): a prospective single-centre study. J Antimicrob Chemother, 2021, 76(1): 233-238.

1. Ababneh MA, Al Domi M, Rababa’h AM. Antimicrobial use and mortality among intensive care unit patients with bloodstream infections: implications for stewardship programs. Heliyon, 2022, 8(8): e10076.
2. Rudd KE, Kissoon N, Limmathurotsakul D, et al. The global burden of sepsis: barriers and potential solutions. Crit Care, 2018, 22(1): 232.
3. Timsit JF, Ruppé E, Barbier F, et al. Bloodstream infections in critically ill patients: an expert statement. Intensive Care Med, 2020, 46(2): 266-284.
4. Andini N, Hu AN, Zhou LM, et al. A “culture” shift: broad bacterial detection, identification, and antimicrobial susceptibility testing directly from whole blood. Clin Chem, 2018, 64(10): 1453-1462.
5. Thomson GK, Jamros K, Snyder JW, et al. Digital imaging for reading of direct rapid antibiotic susceptibility tests from positive blood cultures. Eur J Clin Microbiol Infect Dis, 2021, 40(10): 2105-2112.
6. Jasuja JK, Zimmermann S, Burckhardt I. Applicability and performance of EUCAST’s rapid antimicrobial susceptibility testing (RAST) on primarily sterile body fluids in blood culture bottles in laboratory routine with total lab automation. Eur J Clin Microbiol Infect Dis, 2021, 40(6): 1217-1225.
7. Jasuja JK, Zimmermann S, Burckhardt I. Evaluation of EUCAST rapid antimicrobial susceptibility testing (RAST) for positive blood cultures in clinical practice using a total lab automation. Eur J Clin Microbiol Infect Dis, 2020, 39(7): 1305-1313.
8. Herroelen PH, Heestermans R, Emmerechts K, et al. Validation of rapid antimicrobial susceptibility testing directly from blood cultures using WASPLab^?, including Colibrí^TM and Radian^? in-line carousel. Eur J Clin Microbiol Infect Dis, 2022, 41(5): 733-739.
9. Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing. 32nd ed. CLSI supplement M100. Wayne, PA: Clinical and Laboratory Standards Institute, 2022.
10. European Committee on Antimicrobial Susceptibility Testing (EUCAST). Zone diameter breakpoints for rapid antimicrobial susceptibility testing (RAST) directly from blood culture bottles: Version 4.0. V?xj?, Sweden: EUCAST, 2022.
11. Clinical and Laboratory Standards Institute (CLSI). Verification of commercial microbial identification and antimicrobial susceptibility testingsystems. 1st ed. CLSI guideline M52. Wayne, PA: Clinical and Laboratory Standards Institute, 2015.
12. Bianco G, Boattini M, Comini S, et al. Rapid determination of ceftazidime/avibactam susceptibility of carbapenemase-producing Enterobacterales directly from blood cultures: a comparative evaluation of EUCAST disc diffusion RAST and direct Etest^? RAST. J Antimicrob Chemother, 2022, 77(6): 1670-1675.
13. Berinson B, Olearo F, Both A, et al. EUCAST rapid antimicrobial susceptibility testing (RAST): analytical performance and impact on patient management. J Antimicrob Chemother, 2021, 76(5): 1332-1338.
14. Bin Najeeb MA, Gupta A, Purwar S, et al. Implementing EUCAST rapid antimicrobial susceptibility testing method for sepsis: lessons learned in a tertiary care center. J Infect Dev Ctries, 2021, 15(6): 833-839.
15. Cherkaoui A, Schorderet D, Azam N, et al. Fully automated EUCAST rapid antimicrobial susceptibility testing (RAST) from positive blood cultures: diagnostic accuracy and implementation. J Clin Microbiol, 2022, 60(10): e0089822.
16. Tay?i MR, ?entürk G?, ?ali?kan E, et al. Implementation of the EUCAST rapid antimicrobial susceptibility test (RAST) directly from positive blood culture bottles without the advanced identification systems. J Antimicrob Chemother, 2022, 77(4): 1020-1026.
17. Jonasson E, Matuschek E, Kahlmeter G. The EUCAST rapid disc diffusion method for antimicrobial susceptibility testing directly from positive blood culture bottles. J Antimicrob Chemother, 2020, 75(4): 968-978.
18. Bianco G, Lombardo D, Ricciardelli G, et al. Multicentre evaluation of the EUCAST rapid antimicrobial susceptibility testing (RAST) extending analysis to 16-20 hours reading time. Antibiotics (Basel), 2022, 11(10): 1404.
19. Ekwall-Larson A, Fr?ding I, Mert B, et al. Analytical performance and potential clinical utility of EUCAST rapid antimicrobial susceptibility testing in blood cultures after four hours of incubation. Microbiol Spectr, 2023, 11(2): e0500122.
20. Shan Y, Hu B, Guo W, et al. Evaluation of the EUCAST rapid antimicrobial susceptibility test for Enterobacterales-containing blood cultures in China. J Clin Microbiol, 2022, 60(4): e0255921.
21. Martins A, Wink P, Pereira D, et al. Rapid antimicrobial susceptibility of Enterobacteriaceae by disk diffusion directly from blood culture bottles using the EUCAST RAST breakpoints. J Glob Antimicrob Resist, 2020, 22: 637-642.
22. Valentin T, Koenig E, Prattes J, et al. Implementation of rapid antimicrobial susceptibility testing combined with routine infectious disease bedside consultation in clinical practice (RAST-ID): a prospective single-centre study. J Antimicrob Chemother, 2021, 76(1): 233-238.

West China Medical Journal

Application of total laboratory automation for direct rapid antimicrobial susceptibility testing from Enterobacterales-containing blood cultures

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