Bactec Automated Blood Culture System

Composition, Principle and Procedure

Automated blood culture systems are the primary choice for the detection of pathogens from blood specimens. Instead of manually read broth-based conventional methods, laboratories use continuously monitored, automated blood culture systems for the processing of blood culture specimens nowadays. These systems have increased the yield of blood cultures, reduced the time to organism recovery, and diminished the laboratory technologist’s hands-on time. Several automated blood culture systems are available commercially. Some of the most popular continuous-monitoring blood culture systems are: 

  1. BACTEC (Becton Dickinson and Company),
  2. BacT/ALERT (bioMérieux), and
  3. VersaTREK (Thermo Scientific)

The BD BACTEC™ Automated Blood Culture System utilizes fluorescent technology in detecting the growth of organisms in the blood culture bottles. 

The sample to be tested is inoculated into one or more vials which are inserted into the BD Bactec fluorescent series instrument for incubation and periodic reading. Each vial contains a chemical sensor which can detect an increase in carbon dioxide (CO₂) produced by the growth of microorganisms.

The sensor is monitored by the instrument every ten minutes for an increase in its fluorescence, which is proportional to the amount of CO₂ present. A positive reading indicates the presumptive presence of viable microorganisms in the vial.

Composition of Bactec Blood Culture vials

All the BACTEC culture vials contain soybean-casein digest broth, yeast, amino acids, sugar, vitamins and sodium polyanetholsulfonate. The resin containing vials contains nonionic adsorbing resins and cationic-exchange resins. Sodium polyanetholsulfonate (liquiod) is an anticoagulant and also prevents the killing of bacteria by innate cellular and humoral factors. Resin neutralizes the effect of antibiotics present in the blood and increases the chances of recovery of the pathogen.

The composition of BACTEC™ Plus Aerobic/F culture vials is as follows:

List of Ingredients BD BACTEC™ Plus Aerobic/F
Processed Water 30 mL w/v
Soybean-Casein Digest Broth   3.0%
Yeast Extract   0.25%
Amino Acids   0.05%
Sugar   0.2%
Vitamins 0.025%
Sodium Polyanetholsulfonate (SPS)   0.05%
Antioxidants/Reductants   0.005%
Nonionic Adsorbing Resin   13.4%
Cationic Exchange Resin 0.9%  

Types of Bactec Blood Culture Vials

Bactec Culture vials

Depending on the type of suspected pathogen, several broth formulas are available, including media for the recovery of aerobic and anaerobic bacteria, mycobacteria, and fungi.  No single type is optimal for all microorganisms. BACTEC Plus Aerobic/F, Plus Anaerobic/F, and Peds Plus/F bottles contain resins.

Name of the medium Use
BD BACTEC™ Plus Aerobic medium All-purpose medium capable of supporting the growth of aerobic and facultative organisms.
BD BACTEC™ Plus Anaerobic medium All-purpose medium for common obligate anaerobic and facultative organisms.
BD BACTEC™ Peds Plus media Accommodate small volume and optimally detect common pediatric and non-pediatric organisms
BD BACTEC™ Lytic Anaerobic medium Increase the detection and recovery of anaerobes. Contains detergent to lyse RBCs and WBCs.
BD BACTEC™ Standard Aerobic medium Recovery of bacteria and yeast from the blood.
BD BACTEC™ Standard Anaerobic medium Recovery of anaerobic organisms from the blood.

Procedure

  1. Remove the flip-off cap from the BD BACTEC vial top and inspect the vial for cracks, contamination, excessive cloudiness in medium, and bulging or indented septum. DO NOT USE if any defect is noted.
  2. Before inoculating, swab the septum with alcohol (iodine is NOT recommended).
  3. Aseptically inject or draw directly 8–10 mL of specimen per vial. If the sample volume is decreased recovery will not be as great as with larger volumes (Find more in  Blood Culture: Indications, Timing and Volume)
  4. Place inoculated vials in the BD BACTEC fluorescent series instrument as soon as possible for incubation and monitoring.
  5. Vials entered into the instrument will be automatically tested every ten minutes for the duration of the testing protocol period. Positive vials will be determined by the BD BACTEC fluorescent series instrument and identified as such (see the appropriate BD BACTEC Fluorescent Series Instrument User’s Manual).
  6. Remove the positive vials, subculture in an appropriate solid culture medium (Chocolate Agar, Blood Agar and MacConeky Agar) and Gram-stain. Gram stain will provide a necessary preliminary report which can help physician to take appropriate decision while waiting for the culture report.
  7. If the growth is observed in the solid culture medium, note the colony morphology, do the necessary testing to identify the isolate as well as perform antimicrobial susceptibility test.

Working Principle of BD Bactec Automated Blood Culture System

When microorganisms are present in the test sample inoculated into the BD BACTEC vial, they metabolize nutrients in the culture medium, releasing CO₂ into the medium. A dye in the sensor at the bottom of the culture bottles will react with CO₂. This modulates the amount of light that is absorbed by a fluorescent material in the sensor. The instrument’s photo-detectors measure the level of fluorescence, which corresponds to the amount of CO₂ released by the organisms.

Analysis of the rate and amount of CO₂ increase enables the BD BACTEC fluorescent series instrument to determine if the vial is positive, i.e., that the test sample contains viable organisms.

References and Further Reading

About Acharya Tankeshwar 402 Articles
Hello, thank you for visiting my blog. I am Tankeshwar Acharya. Blogging is my passion. I am working as an Asst. Professor and Microbiologist at Department of Microbiology and Immunology, Patan Academy of Health Sciences, Nepal. If you want me to write about any posts that you found confusing/difficult, please mention in the comments below.

2 Comments

  1. what are the Test that detected food poisonous in the lab ? and which chemical that may kill rate and mice , when you put in the maizes ?

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