Is it possible to perform bacterial identification and antimicrobial susceptibility testing with a positive blood culture bottle for quick diagnosis of bloodstream infections?

Lemos, Tamily Cristina; Cogo, Laura Lúcia; Maestri, Adriane Cheschin; Hadad, Milena; Nogueira, Keite da Silva

doi:10.1590/0037-8682-0311-2017

Cited by 6 publications

(4 citation statements)

References 14 publications

(21 reference statements)

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“…It is based on the observations that mean TTP of +nBCBs is between 8-16 hours and identification of gram-negatives takes ∼4-8 hours in an automated microbial identification system. By direct inoculation of identification cards of various automated systems, studies have shown excellent concordance in identification of gram-negatives [8,23]. By utilizing the widely prevalent automated tools in LMICs and “care-bundle” approach in conjunction, this proposed DSB can act as a primer for improving aBCD in resource limited settings with minimal additional cost.…”

Section: Discussionmentioning

confidence: 99%

A Prospective Study to Improve Automated Blood Culture Diagnostics in Gram-Negative Sepsis by Implementing a “Diagnostic Stewardship Care-Bundle”

Gupta

Siddiqui

Purwar

et al. 2022

Preprint

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ObjectivesWe implemented a diagnostic stewardship care-bundle (DSB): “Sepsis-48” with the aim of evaluating its impact on changes in duration of key steps in automated blood culture diagnostics (aBCD), compliance to care-bundle and turnaround time (TAT).MethodsIn this prospective study, a care-bundle checklist for automated blood culture bottles (BCBs) received from adult intensive care units (AICUs) was implemented between July 2020-June 2021 (intervention period, P2) and compared with a retrospective, pre-intervention period (P1) between March-June 2020. Microbial identification in positive BCBs with gram-negatives (+nBCBs) was enabled by inoculating conventional biochemical tests directly (dID) and direct antimicrobial susceptibility testing (dAST) was done as per EUCAST RAST standard. Clinical reports were issued only if a RAST reportable gram-negative was identified in dID.ResultsWe observed significant reductions in the Loading time (LT) [63.5 (104.5-24.5) vs 32 (55-14) minutes, P<0.001] & time to dID+dAST performance (TTD) [186 (288.25-202.25) vs 115 (180-68) minutes, P=0.0018] in +nBCBs received from AICUs during P2. There was a significant increase in compliance to the bundle targets [LT≤45: 44% vs 66%, P=.006 and TTD≤120: 34% vs 51.7%, P=.03] during P2 (Table 2). Using dID+dAST method, provisional results could be read ∼13 hours earlier than those generated by VITEK®. Similar improvements were also noted for +nBCBs received from other locations.ConclusionsThe “diagnostic stewardship care-bundle” strategy to improve aBCD was successfully implemented leading to significant reductions in duration of targeted steps. Laboratories should implement “diagnostic stewardship care-bundles” as per their needs to improve microbiological diagnostics.

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Section: Discussionmentioning

confidence: 99%

A Prospective Study to Improve Automated Blood Culture Diagnostics in Gram-Negative Sepsis by Implementing a “Diagnostic Stewardship Care-Bundle”

Gupta

Siddiqui

Purwar

et al. 2022

Preprint

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“…However, other novel approaches may help to circumvent this limitation by facilitating timely identification of micro-organisms in such settings. These include direct inoculation of manual biochemicals from pellets obtained by centrifuging the blood-broth mixture from positive BC bottle or even inoculating cards of Automated bacterial identification systems from the pellet [20,21]. Studies have shown excellent correlation for organism identification, particularly in Gram-negatives, where identification cards were directly inoculated from the pellet [20,21].…”

Section: Discussionmentioning

confidence: 99%

Implementing EUCAST rapid antimicrobial susceptibility testing method for sepsis: lessons learned in a tertiary care center

Najeeb¹,

Gupta²,

Purwar³

et al. 2021

J Infect Dev Ctries

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Introduction: We prospectively evaluated EUCAST rapid antimicrobial susceptibility testing methodology for susceptibility testing directly from blood culture bottles in comparison to CLSI disk-diffusion method. Methodology: During May-November 2019, positively flagged blood culture bottles showing Gram-negative micro-organisms were simultaneously processed by rapid antimicrobial susceptibility testing and CLSI methodology. Antibiotics tested were cefotaxime, ceftazidime, piperacillin-tazobactam, imipenem, meropenem, gentamicin, tobramycin and trimethoprim-sulphamethoxazole. Results: Overall, 80 isolates identified as Escherichia coli (n = 24, 30%), Klebsiella pneumoniae (n = 15, 18.7%), Pseudomonas aeruginosa (n = 16, 20%) and Acinetobacter baumannii (n = 25, 31.2%) were included. Categorical agreements of rapid antimicrobial susceptibility testing at 4-, 6- and 8-hour reading times were 88.1% (304/345), 90.8% (425/468) and 92.3% (467/506), respectively. Major Error rates were 14% (21/150), 4.9% (10/206) and 4/236 (1.7%), whereas Very Major Error rates were 1.1% (2/177), 1.3% (3/232) and 3.3% (8/243), respectively. Results categorized as “Area of Technical Uncertainty” were significantly lower at 8-hour {10.2% (39/384) vs 5.2% (28/534), 4- vs 8-hour, p = 0.003, Fischer’s exact test}. Conclusions: Except for a slightly higher Very major error rate, rapid antimicrobial susceptibility testing at 8-hour is equivalent to Disk-diffusion method (CLSI-M100) using CLSI-M52 criteria for equivalence: (Categorical agreement ≥ 90%, Very major error ≤ 1.5% and Major error ≤ 3%). Poor Categorical agreements at all reading times were noted for piperacillin-tazobactam, ciprofloxacin and E. coli. Performance of rapid antimicrobial susceptibility testing methodology in resource limited settings brings unique challenge of identifying micro-organisms within 8 hours. We suggest reading and reporting of results at a single time point using rapid antimicrobial susceptibility testing method i.e. at 8-hour.

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“…To address the high level of antibiotic resistance, a variety of methods have been developed and commercialized such as clinically oriented automated systems including VITEK 2 ® [16] and MALDI-TOF VITEK MS ® systems [17]. Several factors must be considered when developing suitable methods including time saving, cost saving, reliability and robustness of the method, alongside convenience for the end-user.…”

Section: Introductionmentioning

confidence: 99%

3D-Printed Dip Slides Miniaturize Bacterial Identification and Antibiotic Susceptibility Tests Allowing Direct Mastitis Sample Analysis

2022

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The early detection of antimicrobial resistance remains an essential step in the selection and optimization of antibiotic treatments. Phenotypic antibiotic susceptibility testing including the measurement of minimum inhibitory concentration (MIC) remains critical for surveillance and diagnostic testing. Limitations to current testing methods include bulky labware and laborious methods. Furthermore, the requirement of a single strain of bacteria to be isolated from samples prior to antibiotic susceptibility testing delays results. The mixture of bacteria present in a sample may also have an altered resistance profile to the individual strains, and so measuring the susceptibility of the mixtures of organisms found in some samples may be desirable. To enable simultaneous MIC and bacterial species detection in a simple and rapid miniaturized format, a 3D-printed frame was designed for a multi-sample millifluidic dip-slide device that combines panels of identification culture media with a range of antibiotics (Ampicillin, Amoxicillin, Amikacin, Ceftazidime, Cefotaxime, Ofloxacin, Oxytetracycline, Streptomycin, Gentamycin and Imipenem) diluted in Muëller–Hinton Agar. Our proof-of-concept evaluation confirmed that the direct detection of more than one bacterium parallel to measuring MIC in samples is possible, which is validated using reference strains E. coli ATCC 25922, Klebsiella pneumoniae ATCC 13883, Pseudomonas aeruginosa ATCC 10145, and Staphylococcus aureus ATCC 12600 and with mastitis milk samples collected from Reading University Farm. When mixtures were tested, a MIC value was obtained that reflected the most resistant organism present (i.e., highest MIC), suggesting it may be possible to estimate a minimum effective antibiotic concentration for mixtures directly from samples containing multiple pathogens. We conclude that this simple miniaturized approach to the rapid simultaneous identification and antibiotic susceptibility testing may be suitable for directly testing agricultural samples, which is achieved through shrinking conventional tests into a simple “dip-and-incubate” device that can be 3D printed anywhere.

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Is it possible to perform bacterial identification and antimicrobial susceptibility testing with a positive blood culture bottle for quick diagnosis of bloodstream infections?

Abstract: This alternative methodology may assist in the early identification and susceptibility testing of GNB. Further research is necessary to develop appropriate methods for GPC.

Cited by 6 publications

References 14 publications

A Prospective Study to Improve Automated Blood Culture Diagnostics in Gram-Negative Sepsis by Implementing a “Diagnostic Stewardship Care-Bundle”

A Prospective Study to Improve Automated Blood Culture Diagnostics in Gram-Negative Sepsis by Implementing a “Diagnostic Stewardship Care-Bundle”

Implementing EUCAST rapid antimicrobial susceptibility testing method for sepsis: lessons learned in a tertiary care center

3D-Printed Dip Slides Miniaturize Bacterial Identification and Antibiotic Susceptibility Tests Allowing Direct Mastitis Sample Analysis

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