Collection and Inoculation of Blood Specimens for Routine Culture (Bacterial, T.B. or Fungus)


Whenever possible, a peripheral sample is preferred over a sample obtained from a line, unless a line infection is suspected. When drawing multiple blood samples from a line, always draw the blood culture first. Do not discard any blood from the line. Use this initial blood in the line, as this may be the best source if an organism is present.

  • Lab requisitions
  • Name labels 
  • Safety Lok Blood Collection Set/Saf-T E-Z Set 
  • Angel Wing 
  • Blood Culture Bottles* 
  • Skin prep kit (ChloraPrep One-Step Frepp™
  • Tourniquet 
  • Gauze and paper tape 
  • Unsterile examination gloves

*NOTE: Mycobacterial cultures require different containers.

Recommended Testing
  1. Adults: suspected bacteremia – draw two sets of blood culture bottles (aerobic and anaerobic) from two separate venipuncture sites. Maximum of 4 cultures. The aerobic bottle should be inoculated first as there is about 0.5 cc of air in the line of the collection set and sometimes it is difficult to obtain 8-10 cc of blood per bottle (15-20 cc/set). The aerobic bottle is the more critical one to inoculate short samples into. The small lines on the edge of the label indicate approximately 5 cc and there is a fill line denoted on the label. It is important not to underfill or overfill the bottles as this can adversely affect the results.Pediatrics/Neonatal Patient: one aerobic (yellow top) bottle as ordered by the physician. Recommended volume: 1 to 2 cc. of blood per bottle.

  2. Blood Culture for TB: two heparin (green top) or two SPS (yellow top) Vacutainers. No more than once per 24 hour period.

  3. Blood Culture for Fungus: One aerobic (blue top) blood culture bottle.

  1. Confirm identity of patient, making sure that the requisition and labels match the name bracelet.
  2. Explain the procedure to the patient.
  3. Select site for the venipuncture.
  4. Remove cap(s) from bottle(s) and wipe bottle top(s) with 70% alcohol prep pad.
  5. Skin Preparation and Specimen Collection/Inoculation:
    1. ChloraPrep One-Step Frepp™
      1. Locate the vein to be used.
      2. Remove Frepp™ from package. Hold the applicator with ChloraPrep One-Step Frepp™ sponge facing downwards and gently squeeze wings, releasing solution for a controlled flow.
      3. Place sponge on selected venipuncture site and depress once or twice to saturate sponge.
      4. Use a back and forth friction scrub for at least 30 seconds.
      5. Allow area prepared to dry for approximately 30 seconds.
      6. Proceed with collection of blood.
    2. Do not wipe off – if you do, you have then contaminated the site.
  6. Maintain a “hands-off” approach during the prep procedure. If it is necessary to touch the cleansed site, prep the glove finger in the same manner with a fresh prep pad and only palpate above and below the needle insertion site.
  7. a. Peripheral Draw
    1. Using the butterfly Safety E-Z Collection Set and a “male” Angel Wing, perform the venipuncture. Blood should “flash” back into the tubing.
    2. Use the aerobic bottle first and insert blood culture bottles onto adapter. Collect 10 cc. of blood into each bottle.
    Note: Collect blood in the aerobic bottle first as there is about 0.5 cc of air in the line of the butterfly, and in case less than 10 cc is obtained. If less than 10 cc is collected on an adult patient, it should be noted on the requisition as this may produce a false negative test result.
    b. Line Draw
    When drawing from a line, there are two methods:
    1. Directly insert “male” angel wing to the clave connector, OR
    2. Apply syringe to the clave, draw back specimen and then remove syringe and apply “female” angel wing to the syringe.
  8. Put blood into appropriate bottles.
  9. Use an alcohol prep to remove any blood from the gray stopper of the blood culture bottles after the procedure is complete.
  10. Label the bottle(s) with patient name, patient number, time, date, and location of site of specimen, such as "right arm" or "left arm." For line draws, always indicate which line and/or port. Place the label vertically on the bottle and DO NOT place label over the bar code on the blood culture bottle.
    Note: Apply patient label to bottles at bedside after the culture is obtained from the identified patient.
  11. If other blood is required, draw the additional tubes after the blood cultures are obtained.
Aerobic Blue top Bottle 8-10 cc. Blood Peripheral Or Line
Anaerobic Fuchsia top Bottle 8-10 cc. Blood Peripheral Or Line
Fungus Blue top Bottle 8-10 cc. Blood Peripheral Or Line
TB or AFB or Mycobacterium 2 green Vacutainer Tubes (Heparin) 7-10 cc. Blood Peripheral Or Line
Pediatric Blood Culture Yellow top Bottle 1-2 cc. Blood Peripheral Or Line

revised 11/24/17

No Appointment Necessary
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