Standard-of-care blood culture was performed at the University of Iowa Hospitals and Clinics Clinical Microbiology Laboratory. Blood culture bottles (BD Bactec Plus Aerobic/F and Lytic 10/Anaerobic F) were incubated on a Bactec FX unit and time to positivity for each bottle was reported from within the associated BD Epicenter software (Becton Dickinson; Franklin Lakes, New Jersey USA). Bacterial identifications were made with the Bruker BioTyper system [version 4.0.0.1 (5627); Bruker Daltonics, Billerica, MA USA] from subcultures of positive bottles to standard agar media.
Lytic 10 anaerobic f
Manufactured by BD
Sourced in United States
The Lytic/10 Anaerobic/F is a laboratory instrument designed for the rapid and accurate detection and enumeration of anaerobic microorganisms. The device utilizes a specialized culture medium and an automated detection system to provide quantitative results within a short timeframe.
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7 protocols using lytic 10 anaerobic f
1
Standard Blood Culture Protocol
2
Blood Culture Vial Microbial Testing
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The blood culture vials used were the BD BACTEC™ Plus Aerobic/F, Lytic/10 Anaerobic/F, and PEDS Plus/F (Becton, Dickinson and Company, Franklin Lakes, New Jersey, United States of America). First, the blood (if applicable, depending on the experiment) and lock solutions were added to the blood culture vials. It should be noted that, for the experiments without blood, the vials contained a lower overall volume and thereby higher lock concentration (i.e., 30–40 mL media solution depending on the vial type, 1.0 mL spike solution and 1.5 mL lock solution), since the blood (2–8 mL depending on the vial type) was not added. Subsequently, microbial suspensions were added, immediately followed by placement of the vials in the BD BACTEC™ FX instrument for incubation at 35°C for a maximum of five days (bacteria) or seven days (yeasts). Vials detected as positive by the instrument were taken from the machine, the TTP was noted and the content was subcultured on agar to confirm that the blood culture became positive with the micro-organism used for spiking. Vials that remained negative after five or seven days were also subcultured for 48 hours. The vials were confirmed negative if there was no growth detected.
3
Prospective Analysis of Bloodstream Infections
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The study was performed at Grenoble University Hospital, a major teaching hospital (2,100 beds) serving a regional population of 670,000 inhabitants and covering the following specialties: intensive care, general medicine, surgery, geriatric and pediatric medicine, transplants, and oncology. Between November 2016 and November 2017, we prospectively analyzed 187 BSI episodes. Depending on the availability of the RUO cartridges, all first positive blood cultures (BD Bactec Plus Aerobic/F, Lytic/10 Anaerobic/F or Peds Plus/F, Becton Dickinson, Pont de Claix, France) were tested on a given day to avoid selection biases. We included only the first positive blood culture for each BSI episode after microscopic confirmation to exclude false positive bottles and to select the appropriate ePlex Panel(s) to run (one BCID-GP, GN, FP or several panels if Gram stain revealed a polymicrobial sample). We did not exclude suspicion of contaminants. The study design was non-interventional and the results were not reported to the clinicians or used for the management of the patients.
4
UCB Plasma Culture Protocol
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The plasma was detected using the BD BACTEC 9120 (BD Biosciences) Blood Culture System. The outer surface of the Standard/10 Aerobic/F and Lytic/10 Anaerobic/F (BD Biosciences, Sparks, MD, USA) was cleaned with 75% alcohol. The plastic flip cap was removed and the exposed rubber septum was cleaned with an alcohol swab. Then, 3.0–10 mL plasma from UCB was collected for anaerobic bacteria and fungi cultures and another 3.0–10 mL for aerobic bacteria and fungi cultures. The inoculated culture vials were loaded into the instrument, and a temperature of (35 ± 1.5)°C in the racks and (30 ± 1.0)°C within the cabinet was maintained. The plasma was cultured for 7 d. The control assays were carried out by sterile saline for negative control and Escherichia coli ATCC25922 for positive control.
5
Blood Culture Volume Guidelines
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Center guidelines recommend that 2 mL total blood be obtained for blood culture tests sent in an initial sepsis evaluation, with 1 mL inoculated into a single pediatric aerobic (BD Bactec™ Peds Plus/F) and 1 mL into a single anaerobic (BD Bactec™ Lytic/10 Anaerobic/F) bottle. For confirmed infection, 1 mL blood inoculated into an appropriate culture bottle to ensure resolution of bacteremia is sufficient. Blood is obtained after aseptic preparation by venous or arterial phlebotomy or by withdrawing blood from a central catheter. If difficulties arise with obtaining blood for initial evaluation, guidelines recommend that a minimum of 1 mL be cultured, ideally in an aerobic bottle. Therefore, the minimum inoculant considered adequate in this study was 1 mL, in either a single bottle or divided between two culture bottles.
6
Blood Culture Volume Guidelines
7
Blood Culture Collection Protocol
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Whole blood samples (8–10 ml) were collected from the patients and inoculated into each vial of the blood culture set, which includes an aerobic bottle (BD BACTEC Plus Aerobic/F Medium) and an anaerobic bottle (BD BACTEC Lytic/10 Anaerobic/F), according to the hospital policy for taking blood samples for blood culture. Blood culture vials were incubated into the BACTEC blood culture analyser (BD BACTEC FX) within 2 h of taking blood samples.
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