Lab Test

Specimen Collection Manual: Blood for Bacterial Culture


Dearborn, Farmington Hills, Grosse Pointe, Royal Oak, Taylor, Trenton, Troy, & Wayne:

 ADULT: Green (Aerobic) & Orange (Anaerobic) BacT/ALERT Blood Culture Bottle.
 PEDIATRIC: Yellow (Aerobic) BacT/ALERT Blood Culture Bottle.

 Adult bottles: 8-10 mL per bottle is optimal.
 (EXCEPTION: If less than 3 mL, put all the blood in an adult aerobic bottle [green cap].)
 Pediatric bottle: 1-3 mL per bottle is optimal (no minimum).
Blue (Aerobic) BacTALERT® Blood Culture Bottle  Lavendar (Anaerobic) BacTALERT® Blood Culture BottleYellow (Aerobic) Pediatric Blood Culture Bottle
(Green Cap)(Orange Cap) (Yellow Cap)

8-10 mL per bottle is optimal.
(EXCEPTION: If less than 3 mL, put all the blood in
an adult aerobic bottle [green cap].)
1-3 mL per bottle is optimal.
(No Minimum)

Specimen Collection Criteria

Procedure for Routine Blood Culture VENIPUNCTURE COLLECTION:

NOTE: Print the labels for the blood culture bottles immediately before starting the blood collection procedure.

  1. Remove plastic caps from tops of bottles.
  2. Vigorously cleanse the rubber septa of the blood culture bottles and the 3.0 mL discard tube 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®) for an additional 30 seconds.
  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 the venipuncture site using a providone-iodine prep pad 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.
    3. Fill or transfer via syringe at least the first 1 mL of blood into the discard tube and discard the tube.
      NOTE: Blood culture bottles should be standing upright to ensure appropriate volume of blood is added.
    4. For adults, 8-10 mL of blood should be transferred into each blood culture bottle.
    5. For pediatric patients, 1-3 mL should be placed into the pediatric blood culture bottle (no minimum).
  7. It is critical to collect adequate blood volumes per 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. Each set of blood cultures must be drawn from separate venipuncture sites and each venipuncture site must be disinfected.
  10. Send all bottles to the Microbiology laboratory 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 must 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.


  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.
  2. It is recommended that 2-3 sets of blood cultures 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 Medical or Technical Director of Microbiology.
  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 of 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.


  1. BD Bactec™ Procedure Manual, Becton-Dickinson.
  2. Procedure Manual, DuPont Isolator System.
  3. Clinical Microbiology Procedure Handbook, ASM Press, 2016.
  4. ChloraPrep® product literature.


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Microtainer® and Vacutainer® are registered trademarks of Becton, Dickinson and Company.
UroVysion® is a registered trademark of Abbott Laboratories. ThinPrep® is a registered trademark of Hologic, Incorporated.