Generic placeholder image

Infectious Disorders - Drug Targets

Editor-in-Chief

ISSN (Print): 1871-5265
ISSN (Online): 2212-3989

Research Article

A Day Saved is a Life Saved: Direct Antimicrobial Susceptibility Testing from Positively Flagged Blood Culture Bottles and their Concordance with the Routine Method

Author(s): Alisha Aggarwal, Kumar S. Abhishek*, Vibhor Tak, Sukanya Mehrotra, Venkat Goutham Nag and Vidhi Jain

Volume 24, Issue 8, 2024

Published on: 17 April, 2024

Article ID: e170424229017 Pages: 7

DOI: 10.2174/0118715265280460240302165218

conference banner
Abstract

Background: Sepsis is a major health problem worldwide and is associated with high morbidity and mortality with every hour delay in initiation of therapy. A conventional method of blood culture and Antimicrobial Susceptibility Testing (AST) takes around 48-72 hours. Empirical antibiotics need to be administered until the sensitivity report is made available. It has been estimated that 20-50% of the empirical antibiotics are inappropriate, resulting in prolonged hospital stays, adverse effects, and emergence of drug resistance. Additionally, this also puts an extra financial burden on both the patients and healthcare settings. Performing direct Antimicrobial Sensitivity Testing (dAST) is an important tool to reduce turn-around time (TAT) by at least 18-24 hours, thus reducing morbidity and mortality among critically ill patients.

Methods: Direct AST (dAST) was performed from the positively flagged blood culture bottles received between December, 2021 to May, 2022 from Intensive Care Units (ICUs) on Mueller- Hinton Agar (MHA) using four drops of withdrawn blood. dAST was performed for six drugs: Ceftriaxone-30 μg (CTR), Piperacillin/Tazobactam-100/10 μg (PIT), Meropenem-10 μg (MRP), Ciprofloxacin-5 μg (CIP), Aztreonam-30 μg (AT), and Colistin (CL). The zone of inhibition was interpreted as per CLSI M100 ed32, 2022 guidelines. A parallel conventional method was also performed to examine for categorical agreement and disagreement. Identification was carried out using MALDI-TOF MS from the colonies that appeared on the dAST plate on the subsequent day.

Results: A total of 162 positively flagged blood culture bottles were included in the study. The majority of the Gram-negative organisms were from Enterobacterales (n=109), followed by Acinetobacter spp. (n=28) and Pseudomonas aeruginosa (n=25). Out of the 972 isolate-antimicrobial combinations, overall Categorical Agreement (CA) was seen in 936 (96.3%), whereas disagreement was observed in 36 with minor error (mE) in 21 (2.2%), major error (ME) in 7 (0.7%), and very major error (VME) in 8 (0.8%) when compared to the routine method. Categorical agreement (CA) of > 99% was seen in ceftriaxone (CTR) and ciprofloxacin (CIP). In comparison, the lowest CA was observed with meropenem (MRP) at 92%. Colistin dAST was performed using the E-strip method, and the result obtained was highly convincing, with an overall disagreement of only 1.2%.

Conclusion: Rapid dAST from positively flagged blood culture bottles proved to significantly reduce the TAT from the time of sample collection to the first availability of antimicrobial susceptibility report with excellent categorical agreement of > 95% using the conventional disc diffusion method. Results obtained were within the acceptance criteria set by U. S. Food and Drug Administration (FDA) guidelines of > 90% categorical agreement for a new method. We were able to obtain excellent concordance for colistin using the E-strip method. Performing dAST not only saves a “day”, but its proper implementation would save a “life”.

Keywords: Sepsis, blood culture, direct antimicrobial susceptibility testing, turn-around time, categorical agreement, MALDITOF MS.

Graphical Abstract
[1]
Sterling SA, Miller WR, Pryor J, Puskarich MA, Jones AE. The impact of timing of antibiotics on outcomes in severe sepsis and septic shock. Crit Care Med 2015; 43(9): 1907-15.
[http://dx.doi.org/10.1097/CCM.0000000000001142] [PMID: 26121073]
[2]
Shorr AF, Micek ST, Welch EC, Doherty JA, Reichley RM, Kollef MH. Inappropriate antibiotic therapy in Gram-negative sepsis increases hospital length of stay. Crit Care Med 2011; 39(1): 46-51.
[http://dx.doi.org/10.1097/CCM.0b013e3181fa41a7] [PMID: 20890186]
[3]
Schwaber MJ, Navon-Venezia S, Kaye KS, Ben-Ami R, Schwartz D, Carmeli Y. Clinical and economic impact of bacteremia with extended- spectrum-beta-lactamase-producing Enterobacteriaceae. Antimicrob Agents Chemother 2006; 50(4): 1257-62.
[http://dx.doi.org/10.1128/AAC.50.4.1257-1262.2006] [PMID: 16569837]
[4]
Marquet K, Liesenborgs A, Bergs J, Vleugels A, Claes N. Incidence and outcome of inappropriate in-hospital empiric antibiotics for severe infection: A systematic review and meta-analysis. Crit Care 2015; 19(1): 63.
[http://dx.doi.org/10.1186/s13054-015-0795-y] [PMID: 25888181]
[5]
Chaturvedi P, Lamba M, Sharma D, Mamoria VP. Bloodstream infections and antibiotic sensitivity pattern in intensive care unit. Trop Doct 2021; 51(1): 44-8.
[http://dx.doi.org/10.1177/0049475520977043] [PMID: 33283677]
[6]
Kumar A, Roberts D, Wood KE, et al. Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 2006; 34(6): 1589-96.
[http://dx.doi.org/10.1097/01.CCM.0000217961.75225.E9] [PMID: 16625125]
[7]
Vincent JL, Sakr Y, Singer M, et al. Prevalence and outcomes of infection among patients in intensive care units in 2017. JAMA 2020; 323(15): 1478-87.
[http://dx.doi.org/10.1001/jama.2020.2717] [PMID: 32207816]
[8]
Machen A, Drake T, Wang YFW. Same day identification and full panel antimicrobial susceptibility testing of bacteria from positive blood culture bottles made possible by a combined lysis-filtration method with MALDI-TOF VITEK mass spectrometry and the VITEK2 system. PLoS One 2014; 9(2): e87870.
[http://dx.doi.org/10.1371/journal.pone.0087870] [PMID: 24551067]
[9]
Antimicrobial susceptibility test (AST) systems - class II special controls guidance for industry and FDA. FDA 2021. Available from: https://www.fda.gov/medical-devices/guidance-documents-medical-devices-and-radiation-emitting-products/antimicrobial-susceptibility-test-ast-systems-class-ii-special-controls-guidance-industry-and-fda [cited 2023 Jul 29].
[10]
Zilberberg MD, Shorr AF, Micek ST, Vazquez-Guillamet C, Kollef MH. Multi-drug resistance, inappropriate initial antibiotic therapy and mortality in Gram-negative severe sepsis and septic shock: a retrospective cohort study. Crit Care 2014; 18(6): 596.
[http://dx.doi.org/10.1186/s13054-014-0596-8] [PMID: 25412897]
[11]
Quesada MD, Giménez M, Molinos S, et al. Performance of VITEK-2 compact and overnight microscan panels for direct identification and susceptibility testing of Gram-negative bacilli from positive FAN BacT/ALERT blood culture bottles. Clin Microbiol Infect 2010; 16(2): 137-40.
[http://dx.doi.org/10.1111/j.1469-0691.2009.02907.x] [PMID: 19778301]
[12]
Kumar M, Shergill SPS, Tandel K, Sahai K, Gupta RM. Direct antimicrobial susceptibility testing from positive blood culture bottles in laboratories lacking automated antimicrobial susceptibility testing systems. Med J Armed Forces India 2019; 75(4): 450-7.
[http://dx.doi.org/10.1016/j.mjafi.2018.08.010] [PMID: 31719741]
[13]
Lupetti A, Barnini S, Castagna B, Nibbering PH, Campa M. Rapid identification and antimicrobial susceptibility testing of Gram-positive cocci in blood cultures by direct inoculation into the BD Phoenix system. Clin Microbiol Infect 2010; 16(7): 986-91.
[http://dx.doi.org/10.1111/j.1469-0691.2009.03006.x] [PMID: 19681952]
[14]
Rajshekar D, Chaudhari KV, Bhat P, et al. Evaluation of performance of direct disk diffusion test from positively flagged blood culture broth: A large scale study from South India. J Lab Physicians 2019; 11(2): 154-60.
[http://dx.doi.org/10.4103/JLP.JLP_137_18] [PMID: 31160856]
[15]
Infante A, Ortiz de la Tabla V, Martín C, Gázquez G, Buñuel F. Rapid identification and antimicrobial susceptibility testing of Gram-negative rod on positive blood cultures using MicroScan panels. Eur J Clin Microbiol Infect Dis 2021; 40(1): 151-7.
[http://dx.doi.org/10.1007/s10096-020-04014-3] [PMID: 32860091]
[16]
Desai A, Unson E, Weinstein M. Can direct disk diffusion susceptibility testing from positive blood cultures provide earlier results to clinicians? Open Forum Infect Dis 2016; 3 (Suppl. 1): 180.
[http://dx.doi.org/10.1093/ofid/ofw172.47]
[17]
Kong H, Zhang S, Chen X, et al. Rapid identification and antimicrobial susceptibility testing directly from blood cultures of gram-negative and gram-positive isolates. Clin Lab 2013; 59(11+12/2013): 1305-0.
[http://dx.doi.org/10.7754/Clin.Lab.2013.121132] [PMID: 24409665]
[18]
Fonseca e Silva D, Silva-Dias A, Gomes R, et al. Evaluation of rapid colistin susceptibility directly from positive blood cultures using a flow cytometry assay. Int J Antimicrob Agents 2019; 54(6): 820-3.
[http://dx.doi.org/10.1016/j.ijantimicag.2019.08.016] [PMID: 31425793]
[19]
Giordano C, Barnini S. Rapid detection of colistin-resistant Klebsiella pneumoniae using MALDI-TOF MS peak-based assay. J Microbiol Methods 2018; 155: 27-33.
[http://dx.doi.org/10.1016/j.mimet.2018.11.008] [PMID: 30439463]
[20]
Azrad M, Keness Y, Nitzan O, et al. Cheap and rapid in-house method for direct identification of positive blood cultures by MALDI-TOF MS technology. BMC Infect Dis 2019; 19(1): 72.
[http://dx.doi.org/10.1186/s12879-019-3709-9] [PMID: 30658585]
[21]
Barth PO, Roesch EW, Lutz L, de Souza ÂC, Goldani LZ, Pereira DC. Rapid bacterial identification by MALDI-TOF MS directly from blood cultures and rapid susceptibility testing: A simple approach to reduce the turnaround time of blood cultures. Braz J Infect Dis 2023; 27(1): 102721.
[http://dx.doi.org/10.1016/j.bjid.2022.102721] [PMID: 36462577]
[22]
Zengin Canalp H, Bayraktar B. Direct rapid identification from positive blood cultures by MALDI-TOF MS: Specific focus on turnaround times. Microbiol Spectr 2021; 9(3): e01103-21.
[http://dx.doi.org/10.1128/spectrum.01103-21] [PMID: 34908465]

© 2024 Bentham Science Publishers | Privacy Policy