Blood culture identification protocol

A single culture bottle inoculated with a blood sample was incubated in an automated BacT/ALERT 3D system (Biomerieux, France) for a maximum of 5 days. Blood cultures flagged as positive were Gram-stained and subcultured on MacConkey, chocolate, and blood agar plates (Oxide, UK) following standard microbiological techniques. Bacterial isolates were identified based on colony morphology, Gram reaction and biochemical reaction. Yeasts on Gram stained smears from positive blood cultures were identified based on morphology; no attempt was made to identify the fungal species. Staphylococcus aureus was differentiated from coagulase-negative staphylococci (CoNS) based on slide and tube coagulase test methods [27 ]. An isolate was considered a blood culture contaminant in instances when CoNS, viridans streptococci, Micrococcus species, Bacillus species, and Corynebacterium species were detected [14 (link), 28 (link)].

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Shimelis T., Tadesse B.T., W/Gebriel F., Crump J.A., Schierhout G., Dittrich S., Kaldor J.M, & Vaz Nery S. (2020). Aetiology of acute febrile illness among children attending a tertiary hospital in southern Ethiopia. BMC Infectious Diseases, 20, 903.

Publication 2020

BacT/ALERT 3D system

Agar Bacillus Bacterial Blood Blood culture Chocolate Coagulase Corynebacterium Microbiological techniques Micrococcus Oxide Staphylococci Staphylococcus aureus Viridans streptococci Yeasts

Corresponding Organization :

Other organizations : UNSW Sydney, Hawassa University, University of Otago, The George Institute for Global Health, Foundation for Innovative New Diagnostics

Top 5 similar protocols

Variable analysis

independent variables

Inoculation of a single culture bottle with a blood sample

dependent variables

Growth of bacterial isolates detected in blood cultures

control variables

Incubation of the inoculated culture bottle in an automated BacT/ALERT 3D system
Gram-staining and subculturing of blood cultures flagged as positive
Identification of bacterial isolates based on colony morphology, Gram reaction, and biochemical reaction
Identification of yeasts based on morphology
Differentiation of Staphylococcus aureus from coagulase-negative staphylococci (CoNS) using slide and tube coagulase test methods
Consideration of CoNS, viridans streptococci, Micrococcus species, Bacillus species, and Corynebacterium species as blood culture contaminants

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