Lab Test Directory

Specimen Collection Manual: Blood for Bacterial Culture


ADULT: Blue (Aerobic) & Burgandy (Anaerobic) Bactec Blood Culture Bottle.
PEDIATRIC: Pink (Aerobic) Pediatric Blood Culture Bottle.
Adult bottles: 8-10 mL per bottle is optimal.
(EXCEPTION: If less than 3 mL, put all the blood in a Pink Pediatric Bottle.)
Pediatric bottle: 1-3 mL per bottle is optimal (no minimum).
Adult Blue Blood Culture Bottle Adult Lavender Blood Culture Bottle Pediatric Yellow Blood Culture Bottle
                (Blue Collar)            (Lavender Collar)                (Pink Collar)
8-10 mL per bottle is optimal.
(EXCEPTION: If less than 3 mL, put all the blood in a Pink Pediatric Bottle.)
1-3 mL per bottle is optimal.
(No Minimum)
  • A blood culture is one of the most important cultures collected. The purpose of this test is to detect and identify bacteria that might be in a patient's bloodstream.
  • Everyone has bacteria on the skin. When collecting a blood specimen for culture, special precautions must be taken to avoid contaminating the specimen with skin bacteria.
  • To avoid possible bacterial contamination, the venipuncture site must be carefully "scrubbed" with disinfectant prior to performing the phlebotomy. If the blood is contaminated due to poor technique or carelessness, the patient suffers and unnecessary expense is incurred due to prolonged hospitalization and/or unnecessary treatment with antibiotics.

Specimen Collection Criteria

Procedure for Routine Blood Culture VENIPUNCTURE COLLECTION:

  1. Remove plastic caps from tops of bottles.
  2. Vigorously cleanse the rubber septa of the blood culture bottles with 70% isopropyl alcohol wipe and allow to dry. (Do NOT use iodine.)
  3. Thoroughly disinfect venipuncture site by vigorously cleansing for 30 seconds back and forth across the site with a new 70% isopropyl alcohol wipe followed by 2% chlorhexidine antiseptic (ChloraPrep®).
  4. Allow ChloraPrep® to dry completely (at least 30 seconds). Do not wipe the site with guaze. Do not touch the venipuncture site with fingers.
    • NOTE: ChloraPrep® cannot be used on infants less than 2 months old. Scrub site using isopropanol/acetone sepp followed by PVP iodine sepp from center outward in a circular direction. Iodine must dry completely.
  5. Do not wipe the site with gauze. Do not touch the venipuncture site with your fingers. (Do not palpate the vein after skin disinfection unless a sterile glove is worn.)
  6. Collect:
    1. Blood using a butterfly collection system attached to a Vacutainer® holder. Blood is collected directly into a bottle. OR
    2. Blood using a butterfly collection system attached to a syringe. After collection, attach the syringe to a blood transfer device to fill the bottles.
    NOTE: Blood culture bottles should be standing upright to ensure appropriate volume of blood is added.
    1. For adults, 8-10 mL of blood should be transferred into each blood culture bottle.
    2. For pediatric patients, 1-3 mL should be placed into the pediatric blood culture bottle (no minimum).
  7. It is very important to collect adequate blood volumes per bottle.
    • Exception: If less than 3 mL, put all the blood in a pediatric bottle.
  8. Document the exact venipuncture site and exact time of blood collection for each blood culture in the appropriate information system (EPIC or SOFT ID) and on the collection label.
  9. Send bottles to Microbiology immediately.

Inpatient Specimen Preparation


  • Avoid drawing blood through an indwelling intravenous or intra-arterial catheter unless it is needed specifically to evaluate a potential catheter-related infection or if venous access is NOT available. For proper interpretation of catheter cultures, it is recommended that venipuncture blood be obtained for the second blood culture.
    1. Remove plastic tabs from the top of the bottles.
    2. Cleanse the rubber septa with a 70% isopropyl alcohol wipe and allow to dry. Do NOT use iodine.
    3. Clean the port with alcohol prep and allow to dry.
    4. Collect blood into a syringe and fill the bottles as described above.
  • NOTES:
    1. Blood volume is one of the most important variables in detection of a septic episode. Submitting less than the minimum volume of blood may compromise the quality of the blood culture. If less than 3 mL of blood is collected, put all the blood into a pediatric bottle.
    2. It is recommended that 2 sets of blood be obtained within 24 hours and that no blood cultures be obtained after that time unless the patient's condition changes. Ideally, blood should be obtained prior to the administration of systemic antimicrobial agents.
    3. More than 3 blood cultures within 24 hours requires the approval of the Microbiology Medical or Technical Director.
    4. Failure to adequately cleanse the hub or skin is the primary reason that microbial flora from the skin such as coagulase-negative Staphylococcus or Corynebacterium spp. contaminate a blood culture. It is recommended that chlorhexidine be utilized to produce skin antisepsis on all patients except on children less than 2 months old. For hub cleansing, use 70% isopropyl alcohol.
    5. Venipuncture remains the method of choice for obtaining blood for culture. Blood obtained through an indwelling line is twice as likely to yield a contaminant as blood obtained through a properly prepared skin site. If it is absolutely necessary to obtain blood from an indwelling line, it must be paired with another sample obtained by venipuncture to assist in interpretation if there is growth in the culture.
    6. If a fastidious or unusual organism or a mold is suspected, the Laboratory must benotified.
    7. For FUNGAL or ACID-FAST BACILLUS (AFB) Blood Cultures, please see Blood for Fungal Isolation or Blood for Mycobacterial (or AFB) Isolation.


  • Blood Culture bottles should be transported to the Microbiology Laboratory immediately after collection.
  • If bottles are being sent through the pneumatic tube system, it is imperative that no more than 2 bottles (1 set) be inserted into one carrier. Only Pediatric glass bottles must be placed into plastic cones (one bottle per cone). Each cone must be placed in a biohazard bag. Two bottles in cones can be placed in one pneumatic tube - "bottom-to-bottom." Additionally, a foam insert must be added to the pneumatic tube to prevent breakage.
  • Make sure specimen collection label is positioned VERTICAL (PARALLEL) to the bottle length without covering the manufacturer's barcode label or numerical identification. (See phone below.)
  • Do NOT refrigerate inoculated Blood Culture bottles. Bottles may be held at room temperature for short periods of time. Store uninoculated bottles at room temperature. Protect from direct sunlight.
  • Do NOT use bottles beyond the expiration date printed on each bottle.


  1. Bactec Procedure Manual, Becton-Dickinson.
  2. Procedure Manual, DuPont Isolator System.
  3. Clinical Microbiology Procedure Handbook, ASM Press, 2010.
  4. ChloraPrep® product literature.


Name Hospital Phone Pager
Microbiology Laboratory RO (248) 551-8090
Jonathon Hopkins RO (248) 551-3933 (248) 995-2716
Barbara Robinson-Dunn, Ph.D. RO (248) 551-8026 (248) 992-6780
Bobby L. Boyanton, Jr., M.D. RO (248) 551-0130 (248) 992-3262
Microbiology Laboratory Troy (248) 964-6127
Kathleen Hennells Troy (248) 964-8014
Microbiology Laboratory GP (313) 473-1802
Veronica Dostal GP (313) 473-6845

Last Updated